Tricyclic compounds, compositions and methods

ABSTRACT

The present invention is directed to compounds of Formula I: 
     
       
         
         
             
             
         
       
     
     or salt thereof, which are modulators of the glucocorticoid receptor. The compounds and salts of the invention are useful in the treatment of conditions mediated by glucocorticoid receptor activity.

CROSS-REFERENCE TO RELATED PATENTS AND PATENT APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 60/887,981 filed Feb. 2, 2007, which isincorporated herein by reference.

FIELD OF THE INVENTION

The present invention includes compounds that are glucocorticoidreceptor modulators. The present invention also includes compositionsand methods of using compounds and compositions.

BACKGROUND OF THE INVENTION

Glucocorticoid receptor modulators are glucocorticoid receptor ligandsthat are used to treat a variety of conditions because of their powerfulanti-inflammatory, antiproliferative and immunomodulatory activity. J.Miner, et al., Expert Opin. Investig. Drugs (2005) 14(12):1527-1545.

Examples of glucocorticoid receptor modulators include dexamethasone,prednisone, prednisolone, RU-486, and as described in WO 2000/66522 andWO 2004/005229.

Treatment with glucocorticoid receptor modulators is often associatedwith side effects, such as bone loss and osteoporosis.

Identifying a glucocorticoid receptor modulator that is efficacious,potent, and has mitigated side-effects fulfills a medical need.

SUMMARY OF THE INVENTION

In one embodiment, the invention relates to a compound of Formula I:

wherein R¹ is —H or —P(O)(OH)₂; or salt thereof.

In another embodiment, the invention relates to compositions comprisinga compound of Formula I and a carrier. In another embodiment, theinvention relates to a method of contacting a glucocorticoid receptorwith a compound of Formula I. An additional embodiment includes methodsof treating a condition in a subject mediated by glucocorticoid receptoractivity by administering to the subject a compound of Formula I.

DETAILED DESCRIPTION

This detailed description of embodiments is intended only to acquaintothers skilled in the art with the inventions, the principles, and thepractical applications so that others skilled in the art may adapt andapply the inventions in their numerous forms, as they may be best suitedto the requirements of a particular use. These inventions, therefore,are not limited to the embodiments described in this specification, andmay be modified.

A. DEFINITIONS

For the following defined terms, these definitions shall be applied,unless a different definition is given in the claims or elsewhere inthis specification.

The term “carrier” describes an ingredient other than a compound.Carriers may be pharmaceutically acceptable material or vehicle.Examples include liquid or solid filler, diluent, excipient, solvent orencapsulating material.

The phrase “contacting a glucocorticoid receptor” means in vivo, exvivo, or in vitro contact is made with a glucocorticoid receptor andincludes administration of a compound or salt of the present inventionto a subject having a glucocorticoid receptor, as well as, for example,introducing a compound or salt of the invention into a sample containinga cellular, unpurified, or purified preparation containing theglucocorticoid receptor. For example, contacting includes interactionsbetween the compound and the receptor, such as binding.

The phrase “inflammation related condition” includes arthritis,fibromyalgia, ankylosing spondylitis, psoriasis, systemic lupuserythematosus, gout, undifferentiated spondyloarthropy, juvenile-onsetspondyloarthritis, Crohn's disease, ulcerative colitis, irritable bowelsyndrome, inflammatory bowel disease and pain associated with theaforementioned conditions. Specific examples of arthritis includerheumatoid arthritis, osteoarthritis, reactive arthritis, infectiousarthritis, psoriatic arthritis, polyarthritis, juvenile arthritis,juvenile rheumatoid arthritis, juvenile reactive arthritis, and juvenilepsoriatic arthritis.

The term “modulation” or “modulators” includes antagonist, agonist,partial antagonists, and partial agonists.

The term “subject” refers to any animal, including mammals, such asmice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep,horses, primates, or humans.

The term “treating” (and corresponding terms “treat” and “treatment”)includes palliative, restorative, and preventative (“prophylactic”)treating of a subject. The term “palliative treating” refers totreatment that eases or reduces the effect or intensity of a conditionin a subject without curing the condition. The term “preventativetreating” (and the corresponding term “prophylactic treating”) refers totreatment that prevents the occurrence of a condition in a subject. Theterm “restorative treating” (“curative”) refers to treatment that haltsthe progression of, reduces the pathologic manifestations of, orentirely eliminates a condition in a subject. Treating can be done witha therapeutically effective amount of compound, salt or composition thatelicits the biological or medicinal response of a tissue, system orsubject that is being sought by an individual such as a researcher,doctor, veterinarian, or clinician.

B. COMPOUNDS

The present invention comprises, in part, tricyclic compounds of FormulaI. These compounds are useful as glucocorticoid receptor modulators.

The present invention includes a compound of Formula I:

wherein R¹ is —H or —P(O)(OH)₂; or salt thereof.

The present invention includes a compound of Formula II:

wherein R¹ is —H or —P(O)(OH)₂; or salt thereof.

The present invention includes compounds of Formula I or II wherein R¹is —H, or salt thereof.

The present invention includes compounds of Formula I or II wherein R¹is —P(O)(OH)₂, or salt thereof.

The present invention includes(4βS,7R,8αR)-4β-benzyl-7-hydroxy-N-(2-methylpyridin-3-yl)-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamideor salt thereof; and(2R,4αS,10αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10α-octahydrophenanthren-2-yldihydrogen phosphate or salt thereof.

Salts of compounds of the present invention include the acid additionand base salts (including disalts) thereof. In one embodiment, thepresent invention includes a hydrochloride salt of the compound ofFormula I. In another embodiment, the present invention includes acalcium salt of the compound of Formula I. In another embodiment, thepresent invention includes a sodium salt of the compound of Formula I.

Suitable acid addition salts are formed from acids which form non-toxicsalts. Examples include the acetate, aspartate, benzoate, besylate,bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, citrate,edisylate, esylate, formate, fumarate, gluceptate, gluconate,glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride,hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate,maleate, malonate, mesylate, methylsulphate, naphthylate, 2-napsylate,nicotinate, nitrate, orotate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate, stearate,succinate, tartrate, tosylate and trifluoroacetate salts.

Suitable base salts are formed from bas s which form nontoxic salts.Examples include the aluminium, arginine, benzathine, calcium, choline,diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine,potassium, sodium, tromethamine and zinc salts.

For a review on suitable salts, see “Handbook of Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH,Weinheim, Germany, 2002).

A salt may be readily prepared by mixing together solutions of compoundsof the present invention and the desired acid or base, as appropriate.The salt may precipitate from solution and be collected by filtration ormay be recovered by evaporation of the solvent. The degree of ionizationin the salt may vary from completely ionized to almost non-ionized.

The compounds of the present invention may be administered as prodrugs.Thus, certain derivatives which may have little or no pharmacologicalactivity themselves can, when administered into or onto the body, beconverted into compounds of the present invention having the desiredactivity, for example, by hydrolytic cleavage. Such derivatives arereferred to as ‘prodrugs’. Further information on the use of prodrugsmay be found in ‘Pro-drugs as Novel Delivery Systems, Vol. 14, ACSSymposium Series (T Higuchi and W Stella) and ‘Bioreversible Carriers inDrug Design’, Pergamon Press, 1987 (ed. E B Roche, AmericanPharmaceutical Association).

Prodrugs can, for example, be produced by replacing appropriatefunctionalities present in the compounds of the present invention withcertain moieties known to those skilled in the art as ‘pro-moieties’ asdescribed, for example, in “Design of Prodrugs” by H Bundgaard(Elsevier, 1985).

Some examples of such prodrugs include:

(i) where the compound contains an alcohol functionality (—OH), an etherthereof, for example, replacement of the hydrogen with(C₁-C₆)alkanoyloxymethyl; and

(ii) where the compound contains a secondary amino functionality, anamide thereof, for example, replacement of hydrogen withS(C₁-C₁₀)alkanoyl.

Finally, certain compounds of the present invention may themselves actas prodrugs of other compounds of the present invention. For example,certain compounds of Formula I or II could be viewed as a prodrug ofother compounds encompassed by Formula I or II.

All isomers, such as stereoisomers, geometric (cis/trans or Z/E) isomersand tautomeric forms of the compounds or salts are included in the scopeof the present invention, including compounds or salts having more thanone type of isomerism, and mixtures of one or more thereof. For example,the following depicts a compound of Formula I and a tautomer.

Also included are acid addition or base salts wherein the counterion isoptically active, for example, D-lactate or L-lysine, or racemic, forexample, DL-tartrate or DL-arginine.

Isomers may be separated by conventional techniques well known to thoseskilled in the art.

The present invention includes isotopically-labelled compounds of theinvention wherein one or more atoms are replaced by atoms having thesame atomic number, but an atomic mass or mass number different from theatomic mass or mass number usually found in nature.

Isotopically-labeled compounds of the invention can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described in the accompanying Examplesand Preparations using an appropriate isotopically-labelled reagent inplace of the non-labeled reagent previously employed.

For the treatment of the conditions referred to below, the compounds ofthe present invention can be administered. Salts of the compounds of thepresent invention could also be used.

C. Compositions

Compounds or salts of the present invention could be part of acomposition. Compositions can also include one or more compounds orsalts of the present invention. The composition can also include anenantiomeric excess of one or more compounds of the present invention.Other pharmacologically active substances and carriers can be includedin the composition.

One embodiment is a composition comprising a compound of Formula I or asalt thereof. Another embodiment is a composition comprising a compoundof Formula I or a salt thereof and a carrier.

For example, the carrier can be an excipient. The choice of excipientwill to a large extent depend on factors such as the particular mode ofadministration, the effect of the excipient on solubility and stability,and the nature of the dosage form.

The composition can be a solid, a liquid, or both, and may be formulatedwith the compound as a unit-dose composition, for example, a tablet,which can contain from 0.05% to 95% by weight of the active compounds.Compounds or salts of the present invention may be coupled with suitablepolymers as targetable drug carriers.

D. METHODS

The present invention includes a method of contacting a glucocorticoidreceptor with a compound or salt of the present invention.

The present invention also includes a method of treating a conditionmediated by glucocorticoid receptor activity in a subject comprisingadministering to the subject a compound or salt of the presentinvention.

A condition mediated by glucocorticoid receptor activity includes:

a) endocrine disorders, such as primary or secondary adrenocorticalinsufficiency, congenital adrenal hyperplasia, nonsuppurativethyroiditis, and hypercalcemia associated with cancer;

b) rheumatic disorders, such as psoriatic arthritis, rheumatoidarthritis, including juvenile rheumatoid arthritis, ankylosingspondylitis, acute and subacute bursitis, acute nonspecifictenosynovitis, acute gouty arthritis, post-traumatic osteoarthritis,synovitis of osteoarthritis, and epicondylitis;

c) collagen diseases, such as systemic lupus erythematosus, and acuterheumatic carditis;

d) dermatologic conditions, such as pemphigus, bullous dermatitisherpetiformis, severe erythema multiforme (Stevens-Johnson syndrome),exfoliative dermatitis, mycosis fungoides, psoriasis, and seborrheicdermatitis;

e) allergic states, such as seasonal or perennial allergies, allergicrhinitis, bronchial asthma, contact dermatitis, atopic dermatitis, serumsickness, and drug hypersensitivity reactions;

f) ophthalmic diseases and conditions, such as allergic corneal marginalulcers, herpes zoster ophthalmicus, anterior segment inflammation,diffuse posterior uveitis and choroiditis, chronic uveitis, sympatheticophthalmia, allergic conjunctivitis, keratitis, chorioretinitis, opticneuritis, iritis and iridocyclitis;

g) respiratory diseases, such as symptomatic sarcoidosis, Loeffler'ssyndrome, berylliosis, fulminating or disseminated pulmonarytuberculosis, and aspiration pneumonitis;

h) hematologic disorders, such as idiopathic thrombocytopenic purpura,secondary thrombocytopenia, acquired (autoimmune) hemolytic anemia,erythroblastopenia (RBC anemia), and congenital (erythroid) hypoplasticanemia;

i) neoplastic diseases, such as leukemia and lymphoma;

j) edematous states, such as inducing diuresis or emission ofproteinuria in the nephrotic syndrome, without uremia, of the idiopathictype or that due to lupus erythematosus;

k) gastrointestinal diseases, such as ulcerative colitis, regionalenteritis, inflammatory bowel disease, Crohn's disease, gastritis,irritable bowel syndrome;

l) miscellaneous conditions, such as tuberculous meningitis andtrichinosis; and

m) neurological conditions, such as Alzheimer's disease, Parkinson'sdisease, Huntington's disease, amyotrophic lateral sclerosis, spinalcord injury, psychotic major depression, and peripheral neuropathy.

A condition mediated by glucocorticoid receptor activity also includestransplant rejection (e.g., kidney, liver, heart, lung, pancreas (e.g.,islet cells), bone marrow, cornea, small bowel, skin allografts, skinhomografts (such as employed in burn treatment), heart valve xenografts,serum sickness, and graft vs. host disease, autoimmune diseases, such asrheumatoid arthritis, psoriatic arthritis, multiple sclerosis, Type Iand Type II diabetes, juvenile diabetes, obesity, asthma, inflammatorybowel disease (such as Crohn's disease and ulcerative colitis), pyodermagangrenum, lupus (systemic lupus erythematosis), myasthenia gravis,psoriasis, dermatitis, dermatomyositis; eczema, seborrhoea, pulmonaryinflammation, eye uveitis, hepatitis, Grave's disease, Hashimoto'sthyroiditis, autoimmune thyroiditis, Behcet's or Sjorgen's syndrome (dryeyes/mouth), pernicious or immunohaemolytic anaemia, atherosclerosis,Addison's disease (autoimmune disease of the adrenal glands), idiopathicadrenal insufficiency, autoimmune polyglandular disease (also known asautoimmune polyglandular syndrome), glomerulonephritis, scleroderma,morphea, lichen planus, viteligo (depigmentation of the skin), alopeciaareata, autoimmune alopecia, autoimmune hypopituatarism, Guillain-Barresyndrome, and alveolitis; T-cell mediated hypersensitivity diseases,including contact hypersensitivity, delayed-type hypersensitivity,contact dermatitis (including that due to poison ivy), uticaria, skinallergies, respiratory allergies (hayfever, allergic rhinitis) andgluten-sensitive enteropathy (Celiac disease); inflammatory diseasessuch as osteoarthritis, acute pancreatitis, chronic pancreatitis, acuterespiratory distress syndrome, Sezary's syndrome and vascular diseaseswhich have an inflammatory and or a proliferatory component such asrestenosis, stenosis and artherosclerosis.

A condition mediated by glucocorticoid receptor activity also includes:

a) asthma of whatever type, etiology, or pathogenesis, in particularasthma that is a member selected from the group consisting of atopicasthma, non-atopic asthma, allergic asthma, atopic bronchialIgE-mediated asthma, bronchial asthma, essential asthma, true asthma,intrinsic asthma caused by pathophysiologic disturbances, extrinsicasthma caused by environmental factors, essential asthma of unknown orinapparent cause, non-atopic asthma, bronchitic asthma, emphysematousasthma, exercise-induced asthma, allergen induced asthma, cold airinduced asthma, occupational asthma, infective asthma caused bybacterial, fungal, protozoal, or viral infection, non-allergic asthma,incipient asthma, wheezy infant syndrome and bronchiolytis;

b) chronic or acute bronchoconstriction, chronic bronchitis, smallairways obstruction, and emphysema;

c) obstructive or inflammatory airways diseases of whatever type,etiology, or pathogenesis, in particular an obstructive or inflammatoryairways disease that is a member selected from the group consisting ofchronic eosinophilic pneumonia, chronic obstructive pulmonary disease(COPD), COPD that includes chronic bronchitis, pulmonary emphysema ordyspnea associated or not associated with COPD, COPD that ischaracterized by irreversible, progressive airways obstruction, adultrespiratory distress syndrome (ARDS), exacerbation of airwayshyper-reactivity consequent to other drug therapy and airways diseasethat is associated with pulmonary hypertension;

d) bronchitis of whatever type, etiology, or pathogenesis, in particularbronchitis that is a member selected from the group consisting of acutebronchitis, acute laryngotracheal bronchitis, arachidic bronchitis,catarrhal bronchitis, croupus bronchitis, dry bronchitis, infectiousasthmatic bronchitis, productive bronchitis, staphylococcus orstreptococcal bronchitis and vesicular bronchitis, acute lung injury;and

e) bronchiectasis of whatever type, etiology, or pathogenesis, inparticular bronchiectasis that is a member selected from the groupconsisting of cylindric bronchiectasis, sacculated bronchiectasis,fusiform bronchiectasis, capillary bronchiectasis, cysticbronchiectasis, dry bronchiectasis and follicular bronchiectasis.

Another embodiment includes a use of a compound or salt of the presentinvention for use in treating obstructive or inflammatory airwaysdiseases of whatever type, etiology, or pathogenesis, in particular anobstructive or inflammatory airways disease that is a member selectedfrom the group consisting of chronic eosinophilic pneumonia, chronicobstructive pulmonary disease (COPD), COPD that includes chronicbronchitis, pulmonary emphysema or dyspnea associated or not associatedwith COPD, COPD that is characterized by irreversible, progressiveairways obstruction, adult respiratory distress syndrome (ARDS),exacerbation of airways hyper-reactivity consequent to other drugtherapy and airways disease that is associated with pulmonaryhypertension, or asthma of whatever type, etiology, or pathogenesis, inparticular asthma that is a member selected from the group consisting ofatopic asthma, non-atopic asthma, allergic asthma, atopic bronchialIgE-mediated asthma, bronchial asthma, essential asthma, true asthma,intrinsic asthma caused by pathophysiologic disturbances, extrinsicasthma caused by environmental factors, essential asthma of unknown orinapparent-cause, non-atopic asthma, bronchitic asthma, emphysematousasthma, exercise-induced asthma, allergen induced asthma, cold airinduced asthma, occupational asthma, infective asthma caused bybacterial, fungal, protozoal, or viral infection, non-allergic asthma,incipient asthma, wheezy infant syndrome and bronchiolytis.

The present invention includes a method of treating an inflammationbelated condition in a subject comprising administering to the subject acompound or salt of the present invention.

The present invention includes a method of treating conditions such asasthma, dermatitis, inflammatory bowel disease, Alzheimer's disease,psychotic major depression, neuropathy, transplant rejection, multiplesclerosis, chronic uveitis, or chronic obstructive pulmonary disease ina subject comprising administering to the subject a compound or salt ofthe present invention.

The present invention includes a method of treating rheumatoid arthritisin a subject comprising administering to the subject a compound or saltof the present invention.

Rheumatoid arthritis is considered a chronic autoimmune and inflammatorydisease producing inflamed joints, which eventually swell, becomepainful, and experience degradation of cartilage, bone, and ligaments ofthe joint. A result of rheumatoid arthritis is deformity, instability,and stiffness of the joint and scarring within the joint. The jointsdeteriorate at a highly variable rate. Many factors, including geneticpredisposition, may influence the pattern of the disease. People withrheumatoid arthritis may have a mild course, occasional flare-ups withlong periods of remission without disease, or a steadily progressivedisease, which may be slow or rapid. Rheumatoid arthritis may startsuddenly, with many joints becoming inflamed at the same time. Moreoften, it starts subtly, gradually affecting different joints. Usually,the inflammation is symmetric, with joints on both sides of the bodyaffected. Typically, the small joints in the fingers, toes, hands, feet,wrists, elbows, and ankles become inflamed first, followed by the kneesand hips.

Pain associated with rheumatoid arthritis is typically somaticnociceptive joint pain. Swollen wrists can pinch a nerve and result innumbness or tingling due to carpal tunnel syndrome. Cysts may developbehind affected knees, can rupture, causing pain and swelling in thelower legs.

The present invention includes a method of treating dermatitis in asubject comprising administering to the subject a compound or salt ofthe present invention.

The present invention includes a method of treating chronic obstructivepulmonary disease in a subject comprising administering to the subject acompound or salt of the present invention.

The present invention includes a method of treating asthma in a subjectcomprising administering to the subject a compound or salt of thepresent invention.

The present invention includes a method of treating Alzheimer's diseasein a subject comprising administering to the subject a compound or saltof the present invention.

The present invention includes a method of mitigating side effectsassociated with glucocorticoid receptor modulation, comprisingadministering a compound of Formula I to a subject.

The present invention includes a method of mitigating side effectsassociated with prednisolone treatment, comprising administering acompound of Formula I to a subject.

The present invention further comprises methods of treating theaforementioned conditions, diseases, and disorders in a subject or asubject susceptible to having such a condition, by administering to thesubject one or more compounds or salts of the present invention.

In one embodiment, the aforementioned treatment is preventativetreatment.

In another embodiment, the aforementioned treatment is palliativetreatment.

In another embodiment, the aforementioned treatment is restorativetreatment.

E. DOSAGE AND ADMINISTRATION

To select the most appropriate dosage form and route of administrationfor treatment of the proposed indication, the compounds or salts of theinvention can be assessed for their biopharmaceutical properties, suchas solubility and solution stability (across pH), and permeability.

Doses for compounds or salts of the invention range from 0.1 mg to 100mg for oral administration and doses range from 2 mg or less for inhaledadministration. The dose may be administered in single or divided dosesand may fall outside of the typical range given herein.

The dosages are based on an average human subject having a weight ofabout 60 kg to 70 kg. Dosing and dosing regimen depend upon subject anda variety of conditions that may affect dosing (age, sex, body weight,etc.). The physician will readily be able to determine doses forsubjects whose weight falls outside this range, such as infants and theelderly.

Oral Administration

The compounds of the invention and salts thereof may be administeredorally. Oral administration may involve swallowing, so that the compoundor salt enters the gastrointestinal tract, and/or buccal, lingual, orsublingual administration by which the compound or salt enters the bloodstream directly from the mouth.

Formulations suitable for oral administration include solid, semi-solidand liquid systems such as tablets; soft or hard capsules containingmulti- or nano-particulates, liquids, or powders; lozenges (includingliquid-filled); chews; gels; fast dispersing dosage forms; films;ovules; sprays; and buccal/mucoadhesive patches. Further, the compoundor salts of the invention can be administered as a spray drieddispersion.

Liquid formulations include suspensions, solutions, syrups and elixirs.Such formulations may be employed as fillers in soft or hard capsules(made, for example, from gelatin or hydroxypropylmethylcellulose) andtypically comprise a carrier, for example, water, ethanol, polyethyleneglycol, propylene glycol, methylcellulose, or a suitable oil, and one ormore emulsifying agents and/or suspending agents. Liquid formulationsmay also be prepared by the reconstitution of a solid, for example, froma sachet.

The compounds of the invention and salts thereof may also be used infast-dissolving, fast-disintegrating dosage forms such as thosedescribed in Expert Opinion in Therapeutic Patents, 11 (6), 981-986, byLiang and Chen (2001).

For tablet dosage forms, depending on dose, the drug may make up from 1weight % to 80 weight % of the dosage form, more typically from 5 weight% to 60 weight % of the dosage form. In addition to the drug, tabletsgenerally contain a disintegrant. Examples of disintegrants includesodium starch glycolate, sodium carboxymethyl cellulose, calciumcarboxymethyl cellulose, croscarmellose sodium, crospovidone,polyvinylpyrrolidone, methyl cellulose, microcrystalline cellulose,lower alkyl-substituted hydroxypropyl cellulose, starch, pregelatinisedstarch and sodium alginate. Generally, the disintegrant will comprisefrom 1 weight % to 25 weight %, preferably from 5 weight % to 20 weight% of the dosage form.

Binders are generally used to impart cohesive qualities to a tabletformulation. Suitable binders include microcrystalline cellulose,gelatin, sugars, polyethylene glycol, natural and synthetic gums,polyvinylpyrrolidone, pregelatinised starch, hydroxypropyl cellulose andhydroxypropyl methylcellulose. Tablets may also contain diluents, suchas lactose (monohydrate, spray-dried monohydrate, anhydrous and thelike), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystallinecellulose, starch and dibasic calcium phosphate dihydrate.

Tablets may also optionally comprise surface active agents, such assodium lauryl sulfate and polysorbate 80, and glidants such as silicondioxide and talc. When present, surface active agents may comprise from0.2 weight % to 5 weight % of the tablet, and glidants may comprise from0.2 weight % to 1 weight % of the tablet.

Tablets also generally contain lubricants such as magnesium stearate,calcium stearate, zinc stearate, sodium stearyl fumarate, and mixturesof magnesium stearate with sodium lauryl sulphate. Lubricants generallycomprise from 0.25 weight % to 10 weight %, preferably from 0.5 weight %to 3 weight % of the tablet.

Other possible ingredients include anti-oxidants, colourants, flavouringagents, preservatives and taste-masking agents.

Exemplary tablets contain up to about 80% drug, from about 10 weight %to about 90 weight % binder, from about 0 weight % to about 85 weight %diluent, from about 2 weight % to about 10 weight % disintegrant, andfrom about 0.25 weight % to about 10 weight % lubricant.

Solid formulations for oral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

Suitable modified release formulations for the purposes of the inventionare described in U.S. Pat. No. 6,106,864. Details of other suitablerelease technologies such as high energy dispersions and osmotic andcoated particles are to be found in Pharmaceutical Technology On-line,25(2), 1-14, by Verma et al (2001).

Dose ranges for oral administration also include from 0.1 mg to 80 mg,15 mg to 80 mg, 0.1 mg to 25 mg.

Parenteral Administration

The compounds or salts of the invention may also be administereddirectly into the blood stream, into muscle, or into an internal organ.Example 2 could be administered into the blood stream. Suitable meansfor parenteral administration include intravenous, intraarterial,intraperitoneal, intrathecal, intraventricular, intraurethral,intrasternal, intracranial, intramuscular, intrasynovial andsubcutaneous. Suitable devices for parenteral administration includeneedle (including microneedle) injectors, needle-free injectors andinfusion techniques.

Parenteral formulations are typically aqueous solutions which maycontain excipients such as salts, carbohydrates and buffering agents(preferably to a pH of from 3 to 9), but, for some applications, theymay be more suitably formulated as a sterile non-aqueous solution or asa dried form to be used in conjunction with a suitable vehicle such assterile, pyrogen-free water.

The preparation of parenteral formulations under sterile conditions, forexample, by lyophilisation, may readily be accomplished using standardpharmaceutical techniques well known to those skilled in the art.

The solubility of compounds of the present invention and salts thereofused in the preparation of parenteral solutions may be increased by theuse of appropriate formulation techniques, such as the incorporation ofsolubility-enhancing agents.

Formulations for parenteral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease. Thus compounds of the invention may be formulated as asuspension or as a solid, semi-solid, or thixotropic liquid foradministration as an implanted depot providing modified release of theactive compound. Examples of such formulations include drug-coatedstents and semi-solids and suspensions comprising drug-loadedpoly(dl-lactic-coglycolic)acid (PGLA) microspheres.

Topical Administration

The compounds or salts of the invention may also be administeredtopically, (intra)dermally, or transdermally to the skin or mucosa.Example 1 could be administered to the skin. Typical formulations forthis purpose include gels, hydrogels, lotions, solutions, creams,ointments, dusting powders, dressings, foams, films, skin patches,wafers, implants, sponges, fibres, bandages and microemulsions.Liposomes may also be used. Typical carriers include alcohol, water,mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethyleneglycol and propylene glycol. Penetration enhancers may beincorporated—see, for example, J Pharm Sci, 88 (13), 955-958, by Finninand Morgan (October 1999).

Other means of topical administration include delivery byelectroporation, iontophoresis, phonophoresis, sonophoresis andmicroneedle or needle-free (e.g. Powderject™, Bioject™, etc.) injection.

Formulations for topical administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease.

Inhaled/Intranasal Administration

The compounds or salts of the invention can also be administeredintranasally or by inhalation, typically in the form of a dry powder(either alone, as a mixture, for example, in a dry blend with lactose,or as a mixed component particle, for example, mixed with phospholipids,such as phosphatidylcholine) from a dry powder inhaler, as an aerosolspray from a pressurised container, pump, spray, atomiser (preferably anatomiser using electrohydrodynamics to produce a fine mist), ornebuliser, with or without the use of a suitable propellant, such as1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane, or asnasal drops. For intranasal use, the powder may comprise a bioadhesiveagent, for example, chitosan or cyclodextrin.

The pressurized container, pump, spray, atomizer, or nebuliser containsa solution or suspension of the compound(s) of the invention comprising,for example, ethanol, aqueous ethanol, or a suitable alternative agentfor dispersing, solubilising, or extending release of the active, apropellant(s) as solvent and an optional surfactant, such as sorbitantrioleate, oleic acid, or an oligolactic acid. Prior to use in a drypowder or suspension formulation, the drug product is micronised to asize suitable for delivery by inhalation (typically less than 5microns). This may be achieved by any appropriate comminuting method,such as spiral jet milling, fluid bed jet milling, supercritical fluidprocessing to form nanoparticles, high pressure homogenisation, or spraydrying.

Capsules (made, for example, from gelatin orhydroxypropylmethylcellulose), blisters and cartridges for use in aninhaler or insufflator may be formulated to contain a powder mix of thecompound of the invention, a suitable powder base such as lactose orstarch and a performance modifier such as L-leucine, mannitol, ormagnesium stearate. The lactose may be anhydrous or in the form of themonohydrate, preferably the latter. Other suitable excipients includedextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose andtrehalose.

A suitable solution formulation for use in an atomiser usingelectrohydrodynamics may comprise a compound of the present invention,propylene glycol, sterile water, ethanol and sodium chloride.Alternative solvents which may be used instead of propylene glycolinclude glycerol and polyethylene glycol.

Formulations for inhaled/intranasal administration may be formulated tobe immediate and/or modified release using, for example, PGLA. Modifiedrelease formulations include delayed-, sustained-, pulsed-, controlled-,targeted and programmed release.

In the case of dry powder inhalers and aerosols, the dosage unit isdetermined by means of a valve which delivers a metered amount. Units inaccordance with the invention are typically arranged to administer ametered dose or “puff” which may be administered in a single dose or,more usually, as divided doses throughout the day.

Dose ranges for inhaled administration range from 2 mg to less or 1 mgto less.

Combination

The compounds or salts of the invention may be administered incombination with one or more other therapeutic agents, such as a drug.The compound of the present invention or salt thereof may beadministered at the same time or different time as one or more othertherapeutic agents.

For example, “in combination” includes: simultaneous administration of acombination of compound or salt of the invention and a therapeutic agentto a subject, when such components are formulated together into a singledosage form which releases said components at substantially the sametime to said subject; substantially simultaneous administration of acombination of compound or salt of the invention and a therapeutic agentto a subject in need of treatment, when such components are formulatedapart from each other into separate dosage forms which are taken atsubstantially the same time by said subject, whereupon said componentsare released at substantially the same time to said subject; sequentialadministration of a combination of compound or salt of the invention anda therapeutic agent to a subject, when such components are formulatedapart from each other into separate dosage forms which are taken atconsecutive times by said subject with a significant time intervalbetween each administration, whereupon said components are released atsubstantially different times to said subject; and sequentialadministration of such combination of compound or salt of the inventionand a therapeutic agent to a subject, when such components areformulated together into a single dosage form which releases saidcomponents in a controlled manner whereupon they are concurrently,consecutively, and/or overlappingly administered at the same and/ordifferent times by said subject, where each part may be administered byeither the same or different route.

For example, the compounds or salts of the present invention may be usedin combination, partially or completely, in addition to otherantiinflammatories. Suitable antiinflammatories include cyclosporine,zoledronic acid, efalizumab, alefacept, etodolac, lornoxicam, OM-89,vaidecoxib, tocilizumab, abatacept, meloxicam, etanercept, nambumetone,rimexolone, 153Sm-EDTMP, prosorba, imidazole salicylate, oprelvekin,hylauronic acid, naproxen, piroxicam, diacerein, lumericoxib,tacrolimus, aceclofenac, actarit, tenoxicam, rosiglitazone, deflazacort,adalimumab, leflunomide, risedronate sodium, misoprostol and diclofenac,SK-1306X, infliximab, anakinra, celecoxib, diclofenac, etoricoxib andfelbinac, reumacon, golimumab, denosumab, ofatumumab, 10rT1 antibody,pelubiprofen, licofelone, temsirolimus, eculizumab, iguratimod, andprednisone. Other suitable antiinflammatories include CP-481715,ABN-912, MLN-3897, HuMax-IL-15, RA-1, paclitaxel, Org-37663, Org 39141,AED-9056, AMG-108, fontolizumab, pegsunercept, pralnacasan, apilimod,GW-274150, AT-001, 681323 (GSK) K-832, R-1503, ocrelizumab, DE-096,Cpn10, THC+CBD (GW Pharma), 856553 (GSK), ReN-1869, immunoglobulin,mm-093, amelubant, SCIO-469, ABT-874, LenkoVAX, LY-2127399, TRU-015,KC-706, dipyridamole, amoxapinet and dipyridamole, TAK-715, PG 760564,VX-702, prednisolone and dipyridamole, PMX-53, belimumab, prinaberel,CF-101, tgAAV-TNFR:Fc, R-788, prednisolone and SSRI, dexamethasone,CP-690550 and PMI-001.

One of ordinary skill in the art will also appreciate that when usingthe compounds of the invention or salts thereof in the treatment of aspecific disease that the compounds of the invention may be combinedwith various existing therapeutic agents used for that disease.

For example, the compounds or salts of the invention may be combinedwith agents that modulate one or more of the following targets:Cyclooxygenase 2 (prostaglandin endoperoxide-synthase 2); TNF-R (tumornecrosis factor receptor type 1); Cyclooxygenase (Cox 1 and 2;Non-specific); Map Kinase p38 (Non-specific); II1 receptor (type I andII, Non-specific); Arachidonate 5-lipoxygenase; Glucocorticoid receptor(GR); NF-kB; Tumour necrosis factor (TNF-alpha); CCR1 chemokinereceptor; Leukotriene B4 receptor (Non-specific); PDE4(Phosphodiesterase 4; Non-specific); IL6 receptor; Integrin(Non-specific); ADAM-17 (TNF-alpha converting enzyme); ICE (Caspase1/interleukin-1 beta convertase); Prostaglandin Synthesis enzymes(Non-specific); Substance-P receptor (SPR/NK-1 receptor); Prostanoidreceptor (Non-specific); Vascular cell adhesion protein 1 (VCAM 1);MMP-13 (collagenase 3); VEGF Receptor (Non-specific); C5A anaphylatoxinchemotactic receptor, (C5AR); Macrophage migration inhibitory factor(MIF); Purine nucleoside phosphorylase (PNP); Beta 1 interferon; MMP-3(stromelysin 1); CCR2 chemokine receptor; MMP-2 (gelatinase A); Tumornecrosis factor receptor 5 (CD40); CD44 antigen (homing function andIndian blood group system); CCR5 chemokine receptor; Prostaglandin Esynthase; Peroxisome proliferator activated receptor gamma (PPAR-gamma);CXCR4 chemokine receptor; Cathepsin S; Proto-oncogene LCK tyrosinekinase; CXCR3 chemokine receptor; PDGF Receptor; FKBP (12 FK-506); Igsuperfamily CTLA-4; Protein Kinase C (PKC, Non-specific); Integrinalpha-V/beta-5; Cathepsin K; 26S Proteasome; Mineralocorticoid receptor(MR); IkB kinase beta subunit (IKK BETA); Platelet activating factorreceptor (PAF-R); Farnesyl pyrophosphate FPP synthetase; CXCR1 chemokinereceptor; Macrophage colony stimulating factor I receptor (CSF-1R); IL18receptor 1; Adenosine A3 receptor; Granulocyte-macrophagecolony-stimulating factor (GMCSF); SYK tyrosine kinase; CRF receptor(Non-specific); Alpha/Beta tubulin heterodimer; Tyrosine kinase(Non-specific); Amyloid beta; Macrophage colony stimulating factor(MCSF); Tumor necrosis factor ligand superfamily member 11 (receptoractivator of nuclear factor kappa b ligand); Phospholipase(Non-specific); Estrogen receptor (alpha/beta; Non-specific); MMP-9(gelatinase B); Nitric oxide synthase (Non-specific); Inducible NitricOxide Synthase (Non-specific); p53 cellular tumor antigen; Insulin-likegrowth factor 1 (somatomedin C); Nicotinic acetylcholine receptorcomplex; Mu-type opioid receptor (MOR-1); IL11; ERBB/EGF ReceptorTyrosine Kinase (Non-specific); Histamine H2 receptor; Dipeptidylpeptidase IV (DPP IV, CD26); Topoisomerase II; CCR7 chemokine receptor;Bacterial Dihydrofolate Reductase (Non-specific); Beta-tubulin; DNApolymerase (Human, any subunit composition); CCR4 chemokine receptor;CCR3 chemokine receptor; K+ (potassium) Channel (Non-specific);Mitogen-activated protein kinase 14 (MAPK14/P38-alpha); L-type calciumchannels (Non-specific); CCR6 chemokine receptor; PDE3((Phosphodiesterase 3; Non-specific); Cysteine protease (Non-specific);Sodium-dependent noradrenaline transporter (NAT); MAP2Kinase (MEKs;Non-specific); RAF Kinase (Non-specific); Hypoxia-inducible factor 1alpha; NMDA receptor; Estrogen receptor beta (ER-beta); Human DNA;Cholecystokinin type B receptor (CCKB); B1 bradykinin receptor (BK1);P2X purinoceptor 7 (P2X7); Adenosine A2A receptor; Cannabinoid receptor2 (CB2); Sigma opioid receptor; Cannabinoid receptor 1 (CB1); CXCR2chemokine receptor; Complement factor I (C3B/C4B inactivator); ProteinKinase B (RAC-Kinase) (Non-specific); Gamma secretase complex; CRTH2(GPR44); p53-associated gene (MDM2 ubiquitin-protein ligase E3); VIPreceptor (Non-specific); IL1 receptor, type I; IL6 (interferon, beta 2);MMP (Non-specific); Insulin; MMP-2/3/9; Calcitonin/calcitonin-relatedpolypeptide, alpha; Lipoxygenase (Non-specific); vascular endothelialgrowth factor (VEGF); Thrombin; Androgen receptor; Map Kinase(Non-specific); Sex hormone-binding globulin; Chemokine CCL2(MCP1/MCAF); Phospholipase A2; Erythropoietin (EPO); Plasminogen;Gastric proton pump (H+ K+ ATPase); Caspase (Non-specific); FGF receptor(Non-specific); Peroxisome proliferator activated receptor alpha(PPAR-alpha); MIP1a receptor (Non-specific); S100 calcium-bindingprotein (Non-specific); PGE receptor (Non-specific); peptidyl argininedeiminase, type IV; PDGF (a/b) complex; Beta-Lactamase and PBPs (Cellwall biosynthesis); Opioid receptor.(Non-specific);Angiotensin-converting enzyme 1 (ACE1); Urokinase-type plasminogenactivator (UPA); Phosphodiesterase (Non-specific PDE); Progesteronereceptor (PR); 5HT (serotonin) receptor (Non-specific); tumor necrosisfactor (ligand) superfamily, member 5 (CD40 ligand); Thymidylatesynthase; Integrin Alpha4—Paxillin interaction; Integrin alpha-4(VLA-4/CD49D); ERK1; glucose phosphate isomerase (autocrine motilityfactor); Dopamine receptor Non-specific); Chemokine CXCL12 (SDF-1);Microsomal triglyceride transfer protein; Integrin alpha-5/beta-1;Signal transducer and activator of transcription 3 (acute-phase responsefactor); Plasminogen activator inhibitor-1 (PAI-1); Vitamin D3 receptor(VDR/1,25-dihydroxyvitamin D3 receptor); Aromatase complex (P450arom andNADPH-cytochrome P450 reductase); Protein tyrosine phosphatase(Non-specific); 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA)reductase; Integrin beta-1 (Fibronectin receptor beta subunit); Integrinbeta-1/alpha-11; P Selectin (GM P140/granule membrane protein-140);Five-lipoxygenase activating protein (FLAP); H+/K+ ATPase(Non-specific); Na+ (sodium) Channel (Non-specific); Thyroid peroxidase;Brain voltage gated sodium channel alpha-1; Beta-2 adrenergic receptor;BCL1 (Cyclin D1); Thyroid hormone receptor (Non-specific); Vascularendothelial growth factor receptor 2 (VEGFR-2/FLK1); Alpha-V/Beta-6Integrin; Integrin alpha-V (Vitronectin receptor alpha subunit/CD51);SRC Kinase; Pleiotrophin (heparin binding growth factor 8, neuritegrowth-promoting factor 1 ); osteopontin (secreted phosphoprotein 1);toll-like receptor 4 (TLR4); Vanilloid receptor (Non-specific);Pi3Kinase (Non-specific); Poly(ADP-ribose) polymerase (PARP); PPARreceptor (Non-specific); Beta adrenergic receptor (Non-specific);transient receptor potential cation channel, subfamily V, member 1(TRPV1); Topoisomerase l; Histamine H1 receptor; kininogen; IKK Kinase(Non-specific); HIV TAT protein; Toll-like receptor 2; solute carrierfamily 22 (organic cation transporter), member 4 (SLC22A4); RXR receptor(Non-specific); Renin (Angiotensinogenase); Gonadotropin-releasinghormone receptor (GNRH-R); Penicillin Binding Proteins (Cell wallpeptidases); Calmodulin; Mitogen-activated protein kinase 1(MAPK1/ERK2); Calcium channel (Non-specific); Aggrecanase(non-specific); JNK kinase (Non-specific); transthyretin (TTR); CX3CR1receptor; Coagulation factor III (thromboplastin, tissue factor);Sodium-dependent serotonin transporter (5HTT); Colony stimulating factor1 (macrophage); Tissue Transglutaminase (Transglutaminase 2/TGM2);Advanced glycosylation end product-specific receptor; Monoamine oxidase(A and B; Non-specific); Histamine receptor (Non-specific);Sodium-dependent dopamine transporter (DAT); Thrombopoietin(myeloproliferative leukemia virus oncogene ligand, megakaryocyte growthand development factor); Signaling lymphocytic activation molecule;Neutral endopeptidase (NEP/Neprilysin); Endothelin-1 receptor (ETA);Tyrosinase; Mitogen-activated protein kinase 8 (MAPK8/JNK1); IAP(inhibitor of apoptosis) non-specific; Phosphoinositide 3-kinase;Prostaglandin F2-alpha receptor (Prostanoid FP receptor); Human growthhormone; Vasopressin receptor (Non-specific); Mast/stem cell growthfactor receptor (C-KIT); CDK (Non-specific); D4/5HT1a (Dopamine D4receptor, serotonin receptor 1a); Angiopoietin 1 receptor (TIE-2) (TEK);Estrogen receptor alpha (ER-alpha); Epidermal growth factor receptor;Focal Adhesion Kinase (Non-specific); Peripheral benzodiazepine receptor(HPBS); Oxytocinase; Cytosolic phospholipase A2; Endopeptidase(Non-specific); FGFR1 FGF receptor 1; Neurokinin NK1/NK2 receptor;Prolyl 4-hydroxylase complex; Integrin alpha-5 (Fibronectin receptoralpha subunit/VLA-5/CD49E); Muscarinic acetylcholine receptor(Non-specific); Tyrosine-protein kinase JAK3 (JANUS KINASE 3);odc1—ornithine decarboxylase; 5HT3 receptor; Adrenomedullin;Phosphatidylinositol 3-kinase homolog (ataxia-telangiectasia mutatedgene/ATM); Erythropoietin receptor; Connective tissue growth factor;RAC-alpha serine/threonine kinase (Protein kinase B); Toll-like receptor9; Neuronal nitric oxide synthase (NOS1); Kappa-type opioid receptor(KOR-1); Cardiac Na+ channel complex; ERBB-2 receptor protein tyrosinekinase (Tyrosine kinase-type cell surface receptor HER2); Thrombinreceptor (PAR-1); PDE4B (cAMP-specific phosphodiesterase 4B/HSPDE4B);Platelet-derived growth factor beta polypeptide; FKBP-rapamycinassociated protein (FRAP, mTOR); thrombomodulin; HIV Protease(retropepsin); PDE4D (cAMP-specific phosphodiesterase 4D/HSPDE4D);Adenosine kinase; Histone Deacetylase (Non-specific); Prostaglandin E2receptor EP4 subtype (Prostanoid EP4 receptor); Mitogen-activatedprotein kinase kinase 3 (MAP2K3); MMP-12 (metalloelastase); OX40receptor; Non specific human ubiquitin ligase; Sulfonylurea receptor(SUR1 (pancreatic) and SUR2 (cardiac/smooth muscle)); Coagulation factorX (Stuart factor); MAP kinase activated protein kinase 2 (MAPKAPK-2);IgE heavy chain constant region; Dopamine D2+5HT2A receptors;5-hydroxytryptamine 4 receptor (5HT4); Type-1 angiotensin II receptor(AT1); Cytochrome P450 3A4; T-cell cyclophilin (cyclophilin A);Neuromedin K receptor (NKR/NK-3 receptor); Leukotriene B4 receptor;Brutons tyrosine kinase (BTK); Mitogen-activated protein kinase kinase 6(MAP2K6); endoglin; M1/D2/5HT2; Sodium Dependent Noradrenalinetransporter+Dopamine D4 receptor; Mitogen-activated protein kinasekinase 4 (MAP2K4); Heat shock protein Hsp90 A/B; Histidinedecarboxylase; solute carrier family 22 (organic cation transporter),member 5 (SLC22A5); CSK tyrosine kinase; Prolyl endopeptidase; Cysteinylleukotriene receptor (CYSLT1); Nuclear receptor NURR1 (Immediate-earlyresponse protein NOT); Toll-like receptor 3; Proteinase activatedreceptor 2 (PAR-2); Prostacyclin receptor (prostanoid IP receptor);Serine (or cysteine) proteinase inhibitor, clade F (alpha-2 antiplasmin,pigment epithelium derived factor), member 1; Pituitary adenylatecyclase activating polypeptide type i receptor (PACAP-R-1); Tumornecrosis factor (ligand) superfamily, member 10; C-MAF (short form);Acetylcholinesterase (ACHE); Alpha1 adrenergic receptor (Non-specific);GABA A Receptor Bz binding; Lysosphingolipid receptor EDG-1; Mucosaladdressin cell adhesion molecule-1 (MAdCam); alpha-1L adrenergicreceptor; Hepatocyte growth factor receptor (MET Proto-oncogene tyrosinekinase); Muscarinic acetylcholine receptor M3; MEK1; Insulin receptor;GABA receptor (A+B; Non-specific); Phosphatidylinositol 3-kinasecatalytic subunit gamma (P13 kinase gamma); Bone morphogenetic protein 2(BMP2); SKY tyrosine protein kinase receptor (TYRO3) (RSE); discoid indomain receptor family, member 2 (DDR2); KV Voltage-gated potassiumchannel (Non-specific); Sphingosine kinase (Non-specific); High affinitynerve growth factor receptor (TRK-A); Carbonic Anhydrases (all);Thrombopoietin receptor; Vascular endothelial growth factor C;angiotensinogen; ATP-binding cassette, sub-family B (MDR/TAP), member 1(ABCB1) (Multidrug resistance P-glycoprotein (MDR1); Mitogen-activatedprotein kinase kinase 7 (MAP2K7); Muscarinic acetylcholine receptor M1;HIV Reverse transcriptase; PDE5A (cGMP-binding, cGMP-specificphosphodiesterase 5A/HSPDE5A); Alpha adrenergic receptor (Non-specific);Lipoprotein-associated coagulation inhibitor; Carboxypeptidase B2(TAFI); Cholinesterase (Non-specific); B2 bradykinin receptor (BK2);Aldose reductase; Coagulation factor XI (Plasma thromboplastinantecedent); Serine/threonine protein kinase P78; Methionineaminopeptidase 2; Soluble guanylate cyclase (Non-specific); Ribosomalprotein S6 kinase; Metabotropic glutamate receptor 1; Non-receptortyrosine-protein kinase TYK2; Metabotropic glutamate receptor(Non-specific); Vascular endothelial growth factor receptor 3(VEGFR-3/FLT4); Mitogen-activated protein kinase 13 (MAPK13/P38 delta);Fibroblast activation protein (seprase); Corticotropin releasing factorreceptor 1 (CRF1l); Mitogen-activated protein kinase 11 (MAPK11/P38beta); Complement component 5; FL cytokine receptor (FLT3); AMPAreceptor (Glutamate receptors 1-4); Nerve growth factor receptor;Acyl-CoA A: cholesterol acyltransferase 1 (ACAT1); Frizzled-likereceptor smoothened homolog (SMO); G-protein-coupled receptor BONZO(STRL33, CXCR6); IKCa proteins; TGF-beta receptor type II (TGFR-2);HIV-vif protein; 5-hydroxytryptamine 2B receptor (5HT2B); Fatty acidbinding protein (Non-specific); Toll-like receptor 7 (TLR7); Ghrelin;CD36 antigen (collagen type I receptor, thrombospondin receptor);Mitogen-activated protein kinase kinase kinase 3 (MAP3K3/MEKK3);FMLP-related receptor I (FMLP-RI); Sphingosine kinase SPHK1;Histidyl-tRNA synthetase; Mitogen-activated protein kinase 9(MAPK9/JNK2); P2X receptor (Non-specific); Casein kinase I(Non-specific); Sulfotransferases (non specific); Nuclear receptorROR-alpha-1; Catechol O-methyltransferase (COMT); Monoamine oxidase A(MAOA); Gamma-glutamyl hydrolase; Protein kinase C alpha type(PKC-alpha); Mitogen-activated protein kinase 12 (MAPK12/ERK6/P38gamma); Alpha2Delta Calcium Channel; Tissue Factor/Factor VIIa complex;Hookworm neutrophil inhibitory factor; IKr potassium channel; HistamineH4 receptor (JAR3) (PFI-13); 5-hydroxytryptamine 2A receptor (5HT2A);Cholecystokinin type A receptor (CCKA); 11-beta hydroxysteroiddehydrogenase 1; Growth hormone releasing hormone; Nicotinicacetylcholine receptor protein alpha-7; 5HT2 receptor (Non-specific);Sodium/hydrogen exchanger isoform 1 (NHE1); Substance-K receptor(SKR/NK-2 receptor); 5-hydroxytryptamine 1D receptor (5HT1D); 5HT1B/1DReceptors; Sucrase-isomaltase; Beta-3 adrenergic receptor; Calcitoningene-related peptide (GGRP) type 1receptor; Cyclin-dependent kinase 4(CDK4); Alpha-1A adrenergic receptor; P2Y12 platelet ADP receptor;Mitogen-activated protein kinase kinase kinase 5 (MAP3K5) (MEKK5);regulator of G-protein signalling 2; Interleukin-1 receptor-associatedkinase (IRAK); Inorganic pyrophosphatase (ppase); ITK/TSK tyrosinekinase; RAR gamma; AXL tyrosine protein kinase (UFO, GAS6 receptor);Activin receptor-like kinase 1 (ALK-1); Runt-related transcriptionfactor 2; AMP deaminase (Non-specific); CCR8 chemokine receptor; CCR11chemokine receptor; Nociceptin receptor; Insulin-like growth factor Ireceptor; P2Y Receptor (Non-specific); Protein kinase C theta type(NPKC-theta); DNA-methyltransferase non-specific; Phosphorylase Kinase(Non-specific); C3A anaphylatoxin chemotactic receptor (C3AR);sphingosine kinase 2 (SPHK2); Casein kinase II non-specific;Phosphoglycerate kinase 1; UDP-GaI:beatGIcNAc beta1,4-galactosyltransferase 2 (B4GALT2); sapiens solute carrier family 7(cationic amino acid transporter, y+ system), member 5 (SLC7A5); MMP-17(MT-MMP 4); Casein kinase II alpha chain (CK II); Growth arrest specific6 (GAS6); MAP kinase activated protein kinase 3 (MAPKAPK-3); Mitogen andstress-activated protein kinase-1 (MSK1); Prostaglandin D2 synthase (21kD, brain); Pancreatic K+ channel (Non-specific); TGF-beta receptor typeI (TGFR-1/Activin receptor-like kinase 5/ALK-5); Cyclin-dependent kinase2 (CDK2); ACAT (ACAT Enzymes 1 and 2; Non-specific); Delta-type opioidreceptor (DOR-1); 5-hydroxytryptamine 6 receptor (5HT6);5-hydroxytryptamine 1A receptor (5HT1A); 5HT1 receptor (Non-specific);Growth hormone receptor; PDE7 (Phosphodiesterase 7; Non-specific); IgEreceptor (R1 and R2; Non-specific); Cyclin-dependent kinase 1 (CDK1);Farnesyl-protein transferase complex; Prostaglandin D2 receptor(prostanoid DP receptor); Complement C1S component; Histone deacetylase5; dickkopf homolog 1 precursor; P2X purinoceptor 4 (P2X4); Lectin-likeoxidized LDL receptor (LOX-1); Epoxide hydrolase 2 (trans-styrene oxidehydrolase) (soluble epoxide hydrolase) (sEH); dihydrodipicolinatesynthase (dhdps) (DapA); CaM Kinase II Complex; LXR alpha/beta(Non-specific LXR); Second mitochondria-derived activator of caspase;Integrin-linked kinase (ILK); Focal adhesion kinase 2 (FADK 2);Adenosine A2B receptor; WEE1-like protein kinase; Checkpoint kinase(CHK2); Bacterial SecA protein; Nicotinic acetylcholine receptor proteinbeta-2; Mitogen-activated protein kinase kinase kinase 1 (MAP3K1/MEKK1);Protein kinase C zeta type (NPKC-zeta); PDK1(3-phosphoinositide-dependent protein kinase-1); 5-hydroxytryptamine 5Areceptor (5HT5A); Steroid 5-alpha-reductase; Mitogen-activated proteinkinase kinase kinase 8 (MAP3K8/COT); Protein tyrosine phosphatase 1B;P2Y purinoceptor 1 (P2Y1); Alpha-1D adrenergic receptor; Casein kinase Iepsilon (CKI-epsilon); 5-hydroxytryptamine 7 receptor (5HT7);Coagulation factor VII (Eptacog alfa); Pyruvate dehydrogenase kinase(PDHK; Non-specific); PDE7A (cAMP-specific phosphodiesterase7A/HSPDE7A); Glucagon-like peptide 1 receptor (GLP-1R); Influenza rnapolymerase subunit p3 (p2 endonuclease); Viral protease (Non-specific);Topoisomerase IV; Parathyroid hormone receptor (PTH2 receptor); Proteinkinase C beta-I type (PKC-beta-1); Dopamine beta hydroxylase;Galactosyltransferase associated protein kinase P58/GTA; Presynapticprotein SAP97; synovial apoptosis inhibitor 1, synoviolin (SYVN1) (HRD1)(HRD-1); Squalene epoxidase (ERG1); Protein kinase C epsilon type(NPKC-epsilon); Corticotropin releasing factor receptor 2 (CRF2);Intermediate conductance calcium-activated potassium channel (IK1);Nucleoside diphosphate kinase A (NDKA) (NM23-H1); Interleukin-1 receptorassociated kinase 4 (IRAK-4); Glycogen synthase kinase-3 alpha (GSK-3alpha); solute carrier family 22 (organic cation transporter), member 2(SLC22A2); Pyruvate dehydrogenase kinase 1 (PDK1); PAK-alpha kinase(PAK-1); Human 14-3-3 proteins; Isoleucyl-tRNA synthetase;Prenylcysteine carboxyl methyltransferase (PCCMT); CKLF1; and NAALADaseII.

The compounds or salts of the invention may further be administered incombination with one or more agents such as SSRI, matrixmetalloproteinase (MMP) inhibitors, aggrecanase inhibitors, induciblenitric oxide (iNOS) inhibitors, inhibitors of insulin-like growth factor(IGF) expression or activity, inhibitors of fibroblast growth factor(FGF) expression or activity, inhibitors of CD 44 expression oractivity, inhibitors of interleukin (IL) expression or activity,inhibitors of tumor necrosis factor alpha (TNF-alpha) expression oractivity, inhibitors of tumor necrosis factor-inducible protein 6(TSG-6) expression or activity, inhibition of Bikunin expression oractivity, inhibitors of beta-secretase (BACE), inhibitors of PACE-4,inhibition of nuclear receptor rev-ErbA alpha (NR1D1) expression oractivity, inhibition of endothelial differentiation sphingolipidG-protein-coupled receptor 1 (EDG1) expression or activity, inhibitionof proteinase-activated receptor (PAR) expression or activity,inhibition of cartilage-derived retinoic-acid-sensitive protein (CD-RAP)expression or activity, inhibitors of protein kinase C zeta (PKCz),inhibition of resistin expression or activity, inhibition of adisintegrin and metalloproteinase 8 (ADAM8), inhibition of complementcomponent 1 s subcomponent (C1s) expression or activity, inhibition offormyl peptide receptor-like 1 (FPRL1) expression or activity.

Additional examples of agents useful in combination with compounds orsalts of the invention include inhibitors of MMP-2, -3, -9, or -13;inhibitors of aggrecanase-1 or -2; inhibitors of IGF-1 or -2 expressionor activity; inhibitors of FGF-2, -18, or -9 expression or activity; andinhibitors of IL-1, 4 or -6 expression or activity.

Further examples of agents useful in combination with compounds or saltsof the is invention include IGF-1 or -2 antibodies; FGF receptor-2 or -3antagonists, CD 44 antibodies, IL-1, -4 or -6 antibodies, TNF-alphaantibodies; TSG-6 antibodies; bikunin antibodies; NR1D1 antagonists;EDG-1 antagonists; PAR antagonists, CD-RAP antibodies, resistinantibodies, C1s antibodies, and FPRL1 antibodies.

Additional examples of compounds that can be administered with thecompounds or salts of the present invention include: Cyclooxygenase-2(COX-2) selective inhibitors such as celecoxib, rofecoxib, parecoxib,valdecoxib, deracoxib, etoricoxib, and lumiracoxib; opioid analgesicssuch as morphine, hydromorphone, oxymorphone, fentanyl, codeine,dihydrocodeine, oxycodone, hydrocodone, buprenorphine, tramadol, andnalbuphine; nonsteroidal antiinflammatory drugs (NSAIDs) such asaspirin, diclofenac, diflunisal, ibuprofen, fenoprofen, naproxen,nepafenac, and acetaminophen; Phosphodiesterase V inhibitors (PDEV) suchas sildenafil; alpha-2-delta ligands such as gabapentin and pregabalin;and local anaesthetics such as benzocaine, lidocaine, ropivacaine,menthol, camphor and methyl salicylate.

Examples of other types of compounds and classes of compounds that canbe used in combination with the compounds or salts of the presentinvention include: analgesics, barbiturate sedatives; benzodiazepines;Histamine H₁ antagonists having a sedative action; sedatives; skeletalmuscle relaxants; N-methyl-D-aspartic acid (NMDA) receptor antagonists;alpha-adrenergics; tricyclic antidepressants; anticonvulsants such ascarbamazepine; tachykinin (NK) antagonists, particularly NK-3, NK-2 orNK-1 antagonists; muscarinic antagonists; neuroleptics; vanilloidreceptor agonists or antagonists; beta-adrenergics; corticosteroids;Serotonin (5-HT) receptor agonists or antagonists such as a5-HT_(1B,1D), 5-HT_(2A), and 5-HT₃ receptor antagonists; cholinergic(nicotinic) analgesics; cannabinoids; metabotropic glutamate subtype 1receptor (mGluR1) antagonists; serotonin reuptake inhibitors such assertraline; noradrenaline (norepinephrine) reuptake inhibitors such asreboxetine, in particular (S,S)-reboxetine; dual serotonin-noradrenalinereuptake inhibitors such as duloxetine; inducible nitric oxide synthase(iNOS) inhibitors such asS-[2-[(1-iminoethyl)amino]ethyl]-L-homocysteine,S-[2-[(1-iminoethyl)-amino]ethyl]-4,4-dioxo-L-cysteine,S-[2-[(1-iminoethyl)amino]ethyl]-2-methyl-L-cysteine,(2S,5Z)-2-amino-2-methyl-7-[(1-iminoethyl)amino]-5-heptenoic acid,2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)-butyl]thio]-5-chloro-3-pyridinecarbonitrile;2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)butyl]thio]-4-chlorobenzonitrile,(2S,4R)-2-amino-4-[[2-chloro-5-(trifluoromethyl)phenyl]thio]-5-thiazolebutanol,2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)butyl]thio]-6-(trifluoromethyl)-3pyridinecarbonitrile,2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)butyl]thio]-5-chlorobenzonitrile,N-[4-[2-(3-chlorobenzylamino)ethyl]phenyl]thiophene-2-carboxamidine, andguanidinoethyldisulfide; acetylcholinesterase inhibitors; prostaglandinE₂ subtype 4 (EP4) antagonists such asN-[({2-[4-(2-ethyl-4,6-dimethyl-1H-imidazo[4,5-c]pyridin-1-yl)phenyl]ethyl}amino)-carbonyl]-4-methylbenzenesulfonamideor4-[(1S)-1-({[5-chloro-2-(3-fluorophenoxy)pyridin-3-yl]carbonyl}amino)ethyl]benzoicacid; leukotriene B4 antagonists such as1-(3-biphenyl-4-ylmethyl-4-hydroxy-chroman-7-yl)-cyclopentanecarboxylicacid; 5-lipoxygenase inhibitors; and sodium channel blockers.

Combinations with compounds or salts of the present invention alsoinclude analgesics such as acetaminophen, naproxen sodium, ibuprofen,tramadol, trazodone; cyclobenzaprine; aspirin, celecoxib, valdecoxib,indomethacin, and other NSAIDs; antidepressants such as tricyclicantidepressants and selective serotonin reuptake inhibitors, for exampleantidepressants such as amitriptyline, imipramine, nortriptyline,doxepin, fluoxetine, sertraline, and paroxetine; muscle relaxants suchas cyclobenzaprine; sleeping aids such as zolpidem.

Combinations with compounds or salts of the pre-sent invention alsoinclude analgesics such as acetominophen, naproxen sodium, ibuprofen,tramadol, aspirin, celecoxib, valdecoxib, indomethacin, and otherNSAIDs; disease-modifying antirheumatic drugs (DMARDs) such assulfasalazine or methotrexate; corticosteeroids; and tumor necrosisfactor (TNF) blockers such as etanercept and infliximab.

Combinations with compounds or salts of the present invention includetopical corticosteroids; vitamin D analogs such as calcipotriene;anthralin; topical retinoids (i.e., vitamin A derivatives) such asacitretin and tazarotene; clobetasol propionate; methotrexate;azathioprine; cyclosporine; hydroxyurea; and immune-modulating drugssuch as alefacept, efalizumab, and etanercept. Treatment withphototherapy, including psoralen ultraviolet A (psoralen UVA or PUVA)therapy, narrow-band ultraviolet B (UVB) therapy, and combination lighttherapy could be used with compounds or salts of the present inventionand the aforementioned combinations.

Combinations with compounds or salts of the present invention includeNSAIDs such as acetominophen, naproxen sodium, ibuprofen, tramadol,aspirin, celecoxib, valdecoxib, and indomethacin; and corticosteroidssuch as prednisone.

Combinations with compounds or salts of the present invention includeanalgesics such as acetominophen, naproxen sodium, ibuprofen, tramadol,aspirin, celecoxib, valdecoxib, indomethacin, and other NSAIDs;anti-inflammatory drugs; sulfasalazine, mesalamine, balsalazide, andolsalazine; corticosteroids; prednisone; budesonide; immunosuppressantdrugs such as azathioprine, mercaptopurine, TNF blockers such asinfliximab and adalimumab, methotrexate, and cyclosporine; antibioticssuch as metronidazole and ciprofloxacin; anti-diarrheals such asloperamide; laxatives; anticholinergic drugs; antidepressants such astricyclic antidepressants and selective serotonin reuptake inhibitors,for example antidepressants such as amitriptyline, imipramine,nortriptyline, doxepin, fluoxetine, sertraline, and paroxetine;alosetron; and tegaserod.

Compounds or salts of the present invention could also be administeredwith a long acting beta agonist.

Suitable examples of other therapeutic agents which may be used incombination with the compounds or salts of the invention include5-Lipoxygenase (5-LO) inhibitors or 5-lipoxygenase activating protein(FLAP) antagonists, leukotriene antagonists (LTRAs) includingantagonists of LTB₄, LTC₄, LTD₄, and LTE₄, histamine receptorantagonists including H1 and H3 antagonists, α₁- and α₂-adrenoceptoragonist vasoconstrictor sympathomimetic agents for decongestant use,muscarinic M3 receptor antagonists or anticholinergic agents, PDEinhibitors, e.g. PDE3, PDE4 and PDE5 inhibitors, theophylline, sodiumcromoglycate, COX inhibitors both non-selective and selective COX-1 orCOX-2 inhibitors (NSAIDs), oral and inhaled glucocorticosteroids,monoclonal antibodies active against endogenous inflammatory entities,P32 agonists, including long-acting β2 agonists, adhesion moleculeinhibitors including VLA-4 antagonists, Kinin-B₁- and B2-receptorantagonists, Immunosuppressive agents, Inhibitors of matrixmetalloproteases (MMPs), tachykinin NK₁, NK₂ and NK₃ receptorantagonists, elastase inhibitors, adenosine A2a receptor agonists,inhibitors of urokinase, compounds that act on dopamine receptors, e.g.D2 agonists, modulators of the NFκB pathway, e.g. IKK inhibitors,modulators of cytokine signalling pathways such as syk kinase, or JAKkinase inhibitors, agents that can be classed as mucolytics oranti-tussive, and antibiotics.

According to the present invention, compounds or salts of the inventioncan be combined with: H3 antagonists, Muscarinic M3 receptorantagonists, PDE4 inhibitors, glucocorticosteroids, adenosine A2areceptor agonists, β2 agonists, modulators of cytokine signallingpathways such as syk kinase, or, leukotriene antagonists (LTRAs)including antagonists of LTB₄, LTC₄, LTD₄, and LTE₄.

According to the present invention, compounds or salts of the inventioncan also be combined with: glucocorticosteroids, such as inhaledglucocorticosteroids with reduced systemic side effects, includingprednisone, prednisolone, flunisolide, triamcinolone acetonide,beclomethasone dipropionate, budesonide, fluticasone propionate,ciclesonide, and mometasone furoate and mometasone furoate monohydrate;muscarinic M3 receptor antagonists or anticholinergic agents includingin particular ipratropium salts, such as ipratropium bromide, tiotropiumsalts, such as tiotropium bromide, oxitropium salts, such as oxitropiumbromide, perenzepine, and telenzepine, or β2 agonists, such aslong-acting β2 agonists, including salmeterol, formoterol, QAB-149 andCHF-4226.

F. USE IN THE PREPARATION OF A COMPOSITION OR MEDICAMENT

In one embodiment, the present invention comprises methods for thepreparation of a composition or medicament comprising the compounds orsalts of the present invention -for use in treating condition mediatedby glucocorticoid receptor activity.

In another embodiment, the invention comprises the use of one or morecompounds or salts of the present invention in the preparation of acomposition or a medicament for inflammation, inflammation relatedcondition, rheumatoid arthritis, dermatitis, Alzheimer's disease.

The present invention also includes the use of one or more compounds orsalts of the present invention for preparation of a composition or amedicament for treating one or more conditions detailed in the Methodssection.

G. SCHEMES

The compounds of the present invention may be prepared using the methodsillustrated in the general synthetic schemes and experimental proceduresdetailed below. The reactions of the synthetic methods herein arecarried out in suitable solvents which may be readily selected by oneskilled in the art of organic synthesis, said suitable solventsgenerally being any solvent which is substantially nonreactive with thestarting materials (reactants), the intermediates, or products at thetemperatures at which the reactions are carried out. A given reactionmay be carried out in one solvent or a mixture of more than one solvent.Depending on the particular reaction step, suitable solvents for aparticular reaction step may be selected.

Preparation of compounds of the invention can involve the protection anddeprotection of various chemical groups. The need for protection anddeprotection, and the selection of appropriate protecting groups can bereadily determined by one skilled in the art. The chemistry ofprotecting groups can be found, for example, in T. W. Greene and P. G.M. Wuts, Protective Groups in Organic Synthesis, 3rd. Ed., Wiley & Sons,Inc., New York (1999), which is incorporated herein by reference in itsentirety.

Reactions can be monitored according to any suitable method known in theart. For example, product formation can be monitored by spectroscopicmeans, such as nuclear magnetic resonance spectroscopy (e.g., ¹H or ¹³C)infrared spectroscopy, spectrophotometry (e.g., UV-visible), or massspectrometry, or by chromatography such as high performance liquidchromatography (HPLC) or thin layer chromatography.

The starting materials used herein are either commercially available ormay be prepared by routine synthetic methods.

The general synthetic schemes are presented for purposes of illustrationand are not intended to be limiting.

The1(R)-Benzyl-5-bromo-9(S)-hydro-10(R)-hydroxy-10R)-methyl-tricyclo[7.3.1.0^(2.7)]trideca-2,4,6-trien-13-oneof Formula A-8 was prepared using the protocol described in Scheme A,which is generally disclosed in WO 00/66522. Ph depicts Phenyl. Bndepicts Benzyl. Compound A-1 can be purchased (for example, VOUS andRiverside; CAS No. 4133-35-1). Compound A-2 can be prepared as describedin Org. Syn. 1971, 51, 109-112.

The(4βS,7R,8αR)-4β-benzyl-7-hydroxy-N-(2-methylpyridin-3-yl)-7-(trifluoromethyl)-4b,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamidewas prepared as described in Scheme B.

The(2R,4αS,10αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10α-octahydrophenanthren-2-yldihydrogen phosphate of C-3 was prepared as described in Scheme C. Bndepicts benzyl.

The(2R,4αS,10αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10α-octahydrophenanthren-2-yldihydrogen phosphate of C-3 was prepared as described in Scheme D. Bndepicts benzyl. Ph depicts phenyl.

The(2R,4αS,10αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10α-octahydrophenanthren-2-yldihydrogen phosphate of C-3 was prepared as described in Scheme E. Bndepicts benzyl. Ph depicts phenyl.

H. PREPARATIONS AND EXAMPLES

Starting Material A-8 is1(R)-Benzyl-5-bromo-9(S)-hydro-10(R)-hydroxy-10(R)-methyl-tricyclo[7.3.1.0^(2.7)]trideca-2,4,6-trien-13-oneas depicted by the following formula:

Preparation 1:(S)-4α-benzyl-7-bromo-2-ethoxy-3,4,4α,9-tetrahydrophenanthrene

Starting Material A-8 (450 g; 1.17 moles) was dissolved in ethanol (4.5L) at ambient temperature. 21% sodium ethoxide in ethanol (44 mL; 0.12moles) was added and the mixture was heated to reflux for three hours.Once the Starting Material A-8 was consumed, the reaction mixture waschilled to −25° C. Acetyl chloride (250 mL; 3.51 moles) was slowly addedto the mixture while the temperature was maintained near −25° C. Afterthe addition was complete, the mixture was warmed to 0° C. and heldthere until the intermediate enone was consumed. The mixture was scurryat this point. 21% sodium ethoxide in ethanol (1.31 L; 3.51 moles) wasadded to the mixture while the temperature was maintained between −5° C.and 5° C. If the mixture was not basic, more sodium ethoxide was added.The temperature of the mixture was increased to 25° C. and then dilutedwith water (5.9 L). The mixture was filtered and the solid was washedwith water (3×). The title compound (440 g; 85 area %) was obtained as abeige solid. ¹H NMR (DMSO) δ ppm: 1.27 (t, 3H), 1.65 (dt, 1H), 2.06 (d,1H), 2.21 (dd, 1H), 2.49 (m, 1H), 2.65 (m, 2H), 2.89 (m, 2H), 3.85 ) q,2H), 5.45 (m, 2H), 6.44 (d, 2H) 7.06 (m, 2H), 7.25 (d, 1H), 7.33 (dd,1H).

Preparation 2:(S)-4α-benzyl-7-bromo-2,2-(1,2-ethylenedioxy)-1,2,3,4,4α,9-hexahydrophenanthrene

The (S)-4α-benzyl-7-bromo-2-ethoxy-3,4,4α,9-tetrahydrophenanthrene (1270g; 3.2 moles; 85 area %, which may be prepared as described inPreparation 1) was dissolved in toluene (6.45 L). The ethylene glycol(898 mL; 16.1 moles) and p-toluenesulfonic acid (6.1 g; 0.03 moles) wereadded and the reaction heated to reflux. Solvent (1 L) was distilledfrom the mixture and replaced with fresh toluene (1 L). Thisdistillation process was repeated twice more. More p-toluenesulfonicacid (6.1 g) was added each time fresh toluene was added. During thereaction, two intermediates (detected by LC) were formed as thesubstrate was converted into product. The end point of the reaction wasan equilibrium point between the two intermediates and the product. Oncethe endpoint was reached, the mixture was cooled to ambient temperature.The mixture was washed with 0.5 M NaOH (2 L). The phases separatedquickly and both were dark with a small rag layer. The mixture waswashed with water (2 L). The phases separated very slowly. The mixturewas dried by azeotropic distillation. Methanol (4 L) was added to themixture and solvent (4 L) was distilled from the mixture. The methanoladdition and solvent distillation were repeated twice more. Methanol wasadded to the mixture and precipitation occurred a few minutes later.More methanol (4 L) was added to the mixture and then brought to reflux.After 30 minutes, the mixture was cooled to 0° C. The mixture wasfiltered and the solid was washed with chilled methanol (2×2 L). Thesolid was dried in a vacuum oven at 65° C. The title compound (882 g; 98area %) was obtained as a beige solid. ¹H NMR (DMSO) δ ppm: 1.71 (m,2H), 2.06 (m, 2H), 2.31 (dd, 1H), 2.39 (m, 1H), 2.68 (d, 1H), 2.77 (m,1H), 2.86 (dd, 1H), 3.86 (m, 4H), 5.45 (m, 1H), 6.50 (m, 2H), 7.00 (m,4H), 7.37 (dd, 1H), 7.44 (d, 1H).

Preparation 3: (S)-methyl4β-benzyl-7,7-(1,2-ethylenedioxy)-4β,5,6,7,8,10-hexahydrophenanthrene-2-carboxylate

The(S)-4α-benzyl-7-bromo-2,2-(1,2-ethylenedioxy)-1,2,3,4,4α,9-hexahydrophenanthrene(719 g; 1.75 moles, which may be prepared as described in Preparation 2)was dissolved in tetrahydrofuran (7.19 L) and chilled to −70° C. The 1.6M n-butyl lithium in hexane (2270 mL; 2.27 moles) was added at a ratesuch that the temperature was maintained below −60° C. The mixture heldan additional 15 minutes after the addition. Carbon dioxide (108 g; 2.45moles) was added while the temperature was maintained below −60° C. Themixture held an additional 15 minutes after the addition. The mixturewas warmed to ambient temperature. Solvent (7 L) was distilled from themixture at atmospheric pressure. DMF (7 L) was added to the mixture. Themixture was cooled to ambient temperature. Methyl iodide (152 mL; 2.45moles) was added and the mixture was held until the reaction wascompleted (˜1 hour). The mixture was heated to 70° C. and solvent wasdistilled by gradually reducing the pressure to 70 mmHg. Oncedistillation had ceased, the mixture was cooled to room temperature.Water (6.5 L) was slowly added to the mixture to precipitate theproduct. The mixture was filtered and the solid washed with water (3×).The solid was dried on the filter. The crude product (736 g; 74 area %)was obtained as a beige solid. The product was purified bychromatography. 463 g of product was recovered from the chromatography.This material was separated from n-heptane (6130 mL). 394 g of the titlecompound was recovered. Another 70 g of title compound was recoveredfrom the motherliquor by chromatography. ¹H NMR (DMSO) δ ppm: 1.74 (m,2H), 2.10 (m, 2H), 2.33 (dd, 1H), 2.45 (m, 1H), 2.72 (d, 1H), 2.79 (m,1H), 2.94 (dd, 1H), 3.40 (d, 1H), 3.87 (m, 7H), 5.49 (m, 1H), 6.47 (m,2H), 6.93 (m, 2H), 7.01 (m, 1H), 7.42 (d, 1H), 7.64 (d, 1H), 7.79 (dd,1H).

Preparation 4: (4βS,8αR)-methyl4β-benzyl-7,7-(1,2-ethylenedioxy)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxylate

The (S)-methyl4β-benzyl-7,7-(1,2-ethylenedioxy)-4β,5,6,7,8,10-hexahydrophenanthrene-2-carboxylate(201 g; 0.515 moles, which may be prepared as described in Preparation3) and 50 ml of ethylene glycol was dissolved in toluene (2.0 L) in anautoclave. To this was added 10 grams of a 5% Pd/C (dry catalyst). Theautoclave was then sealed and purged with nitrogen (three cycles)followed by hydrogen (three cycles). The reaction was run for 18 hourswith a pressure of 80 psig and temperature of 50° C. HPLC analysis forcompletion and selectivity (typical selectivity's are: 95 to 5, Trans toCis). The suspension was filtered through Celite® to remove the catalystand the toluene solution is concentrated at 50° C., under vacuum, toapproximately 200 ml. While still at 50° C., 1 L of 1-butanol was addedand the solution heated to 60° C., until clear. Upon cooling, theresulting solid title compound was isolated by vacuum filtration (196grams; 97%; Trans to Cis 95.75 to 4.24). ¹H NMR (300 MHz, CDCl₃) δ ppm:7.79 (bs, 1H, Ar—H), 7.47 (d, J=9 Hz, 1H, Ar—H), 7.13-7.05 (cm, 3H,Ar—H), 6.56-6.53 (cm, 2H, Ar—H), 6.43 (d, J=9 Hz, 1H, Ar—H), 4.04-3.93(cm, 4H, 2-CH₂), 3.89 (s, 3H, CH₃), 3.08-3.03 (cm, 3H, CH₂, CH—H), 2.63(d, J=15 Hz, CH—H), 2.22-1.72 (cm, 8H, 4-CH₂), 1.57 (cm, 1H, CH—H).;¹³CNMR (CDCl₃, δ): 167.7, 149.2, 137.7, 136.4, 131.1, 130.5, 127.8,127.7, 127.4, 126.3, 125.5, 108.9, 64.6, 64.5, 52.1, 40.5, 39.8, 38.3,35.8, 31.6, 30.3, 27.9, 24.6.

Preparation 5: (4βS,8αR)-methyl4β-benzyl-7-oxo-4β,5,6,7,8,9,10-octahydrophenanthrene-2-carboxylate

The (4βS,8αR)-methyl4β-benzyl-7,7-(1,2-ethylenedioxy)-4β,5,6,7,8,8α9,10-octahydrophenanthrene-2-carboxylate(150 g, 382 mmol, which may be prepared as described in Preparation 4)was dissolved in dichloromethane (630 ml). Water (270 ml) was added withstirring followed by trifluoroacetic acid (73 ml. 1150 mmol) via dropfunnel over 30 minutes, maintaining the internal temperature below 30°C. After the addition was complete, the reaction was heated at 40° C.for 2 hours. In process check indicated incomplete reaction with around9% (area percent) starting material. The layers were separated and freshwater (270 ml) and trifluoroacetic acid (31 ml) was added. The reactionmixture was heated at 40° C. for 1 hour. This process was continueduntil the starting material was consumed. The organic phase was washedwith 5% aqueous sodium bicarbonate (300 ml), water(300 ml) and driedover MgSO₄ is and concentrated to dryness to give 126.4 g of the titlecompound (representing a 95% yield). ¹H NMR (DMSO) δ ppm: 7.70 (s, 1H),7.37 (d, J=8.4 Hz, 1H), 7.11 (m, 3H), 6.6 (d, J=5.70 Hz, 2H), 6.45 (d,J=8.4 Hz, 1H), 3.80 (s, 3H), 3.80 (m, 2H), 3.04-1.48 (m, 11H).

Preparation 6: (4βS,7R,8αR)-methyl4β-benzyl-7-hydroxy-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxylate

The (4βS,8αR)-methyl4β-benzyl-7-oxo-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxylate(118 g, 0.339 mole, which may be prepared as described in Preparation 5)dissolved in dichloromethane was chilled to −50° C. The solution becameturbid. 1.0 M Tetrabutylammonium fluoride a solution in THF (3.4 ml,0.003 mol) was added with no appreciable temperature change.Trifluorotrimethylsilane (79 ml, 0.51 mol) was added over 20 minuteswith a color change to bright orange to light red in color. The reactionmixture was held at −50° C. for about 2 hours and then allowed to warmto 0° C. Tetrabutylammonium fluoride (340 ml, 0.34 moles) was added veryslowly at 0° C., to the reaction mixture over 45 minutes. An exothermwas observed with gas evolution. The reaction mixture was stirred 10minutes and HPLC analysis indicated complete desilylialation. Water (1L) was added to the reaction mixture and with vigorous stirring andallowed to warm to room temperature. The organic layer was washed withwater (1 L). The organic layer was concentrated and chromatographed toproduce 72 g, 51% of the title compound, with an additional 32 g ofimpure product. ¹H NMR (DMSO) δ ppm: 7.70 (s, 1H), 7.37 (d, J=8.1 Hz,1H), 7.09 (m, 3H), 6.5 (dd, J=1.2, 6.6 Hz, 2H), 6.38 (d, J=8.4 Hz, 1H),3.80 (s, 3H), 3.80 (m, 2H), 3.09-1.21 (m, 13H).

Preparation 7: (4βS,7R,8αR)-methyl4β-benzyl-7-(bis(benzyloxy)phosphoryloxy)-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxylate

The (4βS,7R,8αR)-methyl 4β-benzyl-7-hydroxy-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxylate (5.0 g; 11.9mmol, which may be prepared as in Preparation 6) and 5-methyltetrazole(3.6 g; 43.0 mmol) were mixed together in dichloromethane (50 mL) atambient temperature. Dibenzylphosphoramidite (8.3 mL; 25.1 mmol) wasadded and the mixture was stirred until the reaction was completed (1hour). The mixture was chilled to 0° C. and 30% hydrogen peroxide (10mL) was ad-ded. The reaction was stirred until the oxidation wascompleted (30 minutes). The aqueous phase was separated from the organicphase. The organic phase was washed with 10% sodium meta-bisulfite (50mL). The organic phase was dried with anhydrous magnesium sulfate andconcentrated. The crude product was purified by silica gelchromatography with 15% ethyl acetate in hexanes. The purified titlecompound (8.41 g; 94% yield) was obtained as a colorless oil thatcontained 6% ethyl acetate by weight. ¹H NMR (DMSO): δ 1.31 (t, 1H),1.63-1.92 (m, 3H), 2.05-2.35 (m, 3H), 2.63 (d, 1H), 2.75-3.16 (m, 4H),3.80 (s, 3H), 5.13 (m, 4H), 6.43 (d, 1H), 6.49 (m, 2H), 7.04-7.17 (m,3H), 7.33-7.42 (m, 12H), 7.71 (d, 1H).

Preparation 8: dibenzyl(2R,4αS,10αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10α-octahydrophenanthren-2-ylphosphate

The (4βS,7R,8αR)-methyl4β-benzyl-7-(bis(benzyloxy)phosphoryloxy)-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxylate(7.9 g; 11.6 mmol, which may be prepared as in Preparation 7) and3-amino-2-picoline (1.3 g; 12.2 mmol) were mixed together intetrahydrofuran (80 mL) and chilled to 0° C. The 1 M solution of lithiumbis(trimethylsilyl)amide in tetrahydrofuran (24 mL; 24.4 mmol) was addedwhile maintaining the temperature below 10° C. The mixture was stirredfor 30 minutes. Water (50 mL) was added to the reaction mixture. Themixture was extracted with ethyl acetate. The organic extract was washedwith water. The organic phase was dried with anhydrous magnesium sulfateand concentrated. The crude product was purified by silica gelchromatography with 70% ethyl acetate in hexanes. The purified titlecompound (6.79 g; 68% yield) was obtained as a yellow gum that contained6% ethyl acetate by weight. ¹H NMR (DMSO): δ 1.33 ft, 1H), 1.66-1.93 (m,3H), 2.08-2.34 (m, 3H), 2.41 (s, 3H), 2.68 (d, 1H), 2.76-3.19 (m, 4H),5.14 (m, 4H), 6.47 (d, 1H), 6.56 (m, 2H), 7.07-7.19 (m, 3H), 7.20-7.53(m, 12H), 7.71 (d, 1H), 7.76 (s, 1H), 8.32 (d, 1H), 9.93 (s, 1H).

Example 1(4βS,7R,8αR)-4β-benzyl-7-hydroxy-N-(2-methylpyridin-3-yl)-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide

The (4βS,7R,8αR)-methyl4β-benzyl-7-hydroxy-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxylate(10 g; 23.9 mmol, which may be prepared as described in Preparation 6),and 3-amino-2-picoline (2.71 g; 25.1 mmol) were dissolved in toluene(200 mL). The 1M lithium bis(trimethylsilyl)amide in tetrahydrofuran(74.1 mL; 74.1 mmol) was added at a rate such that the temperature wasmaintained below 35° C. There was a mild exotherm and a solidprecipitated during the addition. The mixture was held an additional 30minutes after the addition. Water (250 mL) was added to the mixture.There was a mild exotherm and the solid dissolved. Ethyl acetate (50 mL)was added to the mixture to ensure the product did not precipitate.Stirring was stopped to allow the phases to separate. The aqueous phasewas removed. The organic phase was washed with water (250 mL). Solvent(230 mL) was distilled at atmospheric pressure from the organic phase.The mixture was cooled to ambient temperature. The mixture was filteredand the solid was washed with toluene (2 times) followed by heptane (2times). The solid was dried in a vacuum oven at 70° C. The titlecompound of the present example (10 g) was obtained as a beige solid. ¹HNMR (DMSO) δ ppm: 1.32 (m, 1H), 1.82 (m, 4H), 2.10 (m, 4H), 2.41 (s,3H), 2.68 (d, 1H), 3.08 (m, 3H), 6.00 (s, 1H), 6.43 (d, 1H), 6.59 (m,2H), 7.12 (m, 3H), 7.25 (dd, 1H), 7.44 (dd, 1H), 7.71 (dd, 1H), 7.75 (d,1H), 8.31 (dd, 1H), 9.91 (s, 1H).

Example 2(2R,4αS,1αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10αR-octahydrophenanthren-2-yldihydrogen phosphate

The dibenzyl(2R,4αS,10αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10α-octahydrophenanthren-2-ylphosphate (6 g; 7.9 mmol, which may be prepared as described inPreparation 8) was dissolved in methanol (12.0 mL). 5% palladium oncarbon (63% water) (1.3 g; 0.4 mmol) was added to the mixture. Themixture was treated with hydrogen (50 psi) at room temperature. Thereaction stalled with 12% of the monobenzylic intermediate remaining.The mixture was filtered through a pad of Celite®. Fresh catalyst (1.3g) was added to the solution and resubmitted to the hydrogenationconditions. Once the reaction was completed, the mixture was filteredthrough a pad of Celite®. The solution was concentrated to about 60 mLby distillation and not by using a rotary evaporator. During thedistillation a white solid precipitated. The mixture was cooled toambient temperature. The mixture was filtered and the solid washed withmethanol. The solid was dried in a vacuum oven at 70° C. The compound ofthe present example (3.36 g; 75% yield) was obtained as a white solidand had an LC purity of 98 area %. ¹H NMR (DMSO): δ 1.33 (t, 1H),1.69-1.98 (m, 3H), 2.07-2.29 (m, 3H), 2.42 (s, 3H), 2.61-2.80 (m, 2H),2.93-3.19 (m, 3H), 3.30 (d, 1H), 6.50 (d, 1H), 6.64 (m, 2H), 7.08-7.20(m, 3H), 7.29 (dd, 1H), 7.48 (dd, 1H), 7.75 (dd, 2H), 8.33 (dd, 1H),9.96 (s, 1H).

I. Biological Data

For the following descriptions, the comparators are tricyclic compounds(see e.g., WO 2000/66522). The example and comparators compounds wereprepared at Pfizer. Prednisolone was used as a clinically relevantcomparator (P-6004; Sigma-Aldrich, St. Louis).

Comparator A is(4β,7S,8αR)-4β-benzyl-7-hydroxy-N-((2-methylpyridin-3-yl)methyl)-7-(3,3,3-trifluoropropyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator B is(4βS,7R,8αR)-4β-benzyl-N-(3,5-dimethylpyrazin-2-yl)-7-hydroxy-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator C is(4βS,7S,8αR)-4β-benzyl-7-hydroxy-N-(2-methylpyridin-3-yl)-7-(3,3,3-trifluoropropyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator D is(4βS,7R,8αR)-4β-benzyl-7-hydroxy-N-(4-methylpyridin-3-yl)-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator E is(4βS,7R,8αS)-4β-benzyl-7-hydroxy-N-(2-methylpyridin-3-yl)-10-oxo-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator F is(4βS,7R,8αR,10R)-4β-benzyl-7,10-dihydroxy-N-(2-methylpyridin-3-yl)-7-(trifluoromethyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator G is:

Comparator H is(4βS,7R,8αR)-4β-benzyl-7-(difluoromethyl)-7-hydroxy-N-(2-methylpyridin-3-yl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator I is(4βS,7R,8αS)-4β-benzyl-7-hydroxy-N-(2-methylpyridin-3-yl)-7-(trifluoromethyl)-4β,5,6,7,8,8α-hexahydrophenanthrene-2-carboxamide,having the following structure:

Comparator J is (4βS,7S,8αR)-4β-benzyl-N-(2,4-dimethylpyrimidin-5-yl)-7-hydroxy-7-(3,3,3-trifluoropropyl)-4β,5,6,7,8,8α,9,10-octahydrophenanthrene-2-carboxamide,having the following structure:

Comparator K is(2R,4αS,10αR)-4α-benzyl-7-((2-methylpyridin-3-yl)carbamoyl)-2-(trifluoromethyl)-1,2,3,4,4α,9,10,10α-octahydrophenanthren-2-ylisobutyl carbonate, having the following structure:

Conversion of Example 2 into Example 1

The Caco-2 cell monolayer is an in vitro tissue culture model of theintestinal epithelium. These cells are of human colonic origin andbecome polarized, fully differentiated enterocytes in 2-3 weeks. Oncedifferentiated, these cells have tight junctions and express variousbiochemical processes such as active efflux transporters includingP-glycoprotein (P-gp). With this model, it is possible to determine theapparent permeability (P_(app)) of a compound across the polarizedCaco-2 cell monolayer.

An A→B Assay is performed with Caco-2 cell monolayers to determinecompound P_(app) from A chamber to B chamber. This P_(app) isrepresentative of luminal (gut) to serosal (blood) compound transportacross intestinal epithelium that may be seen during intestinalabsorption.

Example 2 did not significantly traverse a Caco-2 cell monolayer (A→B,P_(app)=1.15e⁻⁶ cm/sec), whereas application of Example 2 to the apicalcompartment resulted in a significant elevation of Example 1 in both theapical and basolateral compartments. The data indicates a mechanisminvolving dephosphorylation of Example 2 to Example 1 via membrane-boundalkaline phosphatases located in the intestinal epithelia, followed byabsorption of Example 1 across the Caco-2 cell monolayer (A→B,P_(app)=37.5e⁻⁶ cm/sec).

Oral dosing of Example 2 (30 and 200 mg/kg) in portal vein-cannulatedrats resulted in the detection of Example 1, but not Example 2, inportal vein plasma samples over a four-hour time period. These resultsindicated the occurrence of intestinal first-pass hydrolysis of Example2 to Example 1 and selective intestinal absorption of Example 1.

Example 2 demonstrates enhanced solubility and intrinsic dissolution,which results in an improved oral absorption profile in rats viaincreased exposure (1.61 μg·hr/mL [vs. 0.46 μg·hr/mL for Example 1]) andC_(max) (0.59 μg/mL [vs. 0.13 μg/mL for Example 1]), as well asdecreased time to C_(max) (0.8 hr [vs. 1.5 hr for Example 1]) in dogs.The bioavailability of Example 2 was improved compared with Example 1 inrats is (F=59% for Example 2; F=17% for Example 1).

In Vitro Data

TNFα GRFP IC₅₀ IL-6 IL-6 IC₅₀ TNFα Name (nM) IC₅₀ (nM) % Inhibition (nM)% Inhibition Example 1 HCL 1.31 .400 76.9 92.1 (28.8)^(a) 77.1 (62)^(a)salt Example 1 free .360 86.1 base Example 2 79.0 (42)^(a) 60 (17)^(a)60.2 (80)^(a)  Comparator A 7.10 >36.4 59.9 >1000  35 Comparator B 0.354 75.8 Comparator C 1.22 1.1 82.6 Comparator D 2.06 1.3 79.8 ComparatorE 1.18 1.1 79.7 Comparator F 1.9 7 83.3 Comparator G 2.13 2.1 80.1Comparator H 8.03 4.3 64.9 Comparator I 9.12 2.8 67.5 Comparator J 4.58291 55 Comparator K 895 Prednisolone 0.526 4.2 (4.6)^(a) 102 (100)^(a)14.9 (15.6)^(a) 100 ^(a)indicates additional results that were found.

GRFP: Glucocorticoid Receptor Binding

The glucocorticoid receptor fluorescence polarization ligand binding(GRFP) assay is used to evaluate direct binding of testing compounds tofull-length glucocorticoid (OR) protein. Reagents for this assay arepurchased from Invitrogen in a test kit. A fluorescent labeled GR ligandis used as a fluorescent tracer and test compounds compete with thefluorescent tracer for OR binding. The change in polarization value inthe presence of test compounds is due to binding of test compounds to GRand is used to determine IC₅₀ and relative binding affinity of testcompounds for GR.

IL-6 IC₅₀ and % Inhibition

Human A549 lung epithelial cells (American Type Culture Collection,Rockville, Md.) were cultured in Kaighn's F-1 2K medium withpenicillin-streptomycin (10 U/mL) and 10% heat-inactivated fetal bovineserum (all from Invitrogen, Grand Island, N.Y.). A549 cells were platedat a density of 30,000 cells/well in 96-well plates and incubatedovernight at 37° C. and with 5% CO₂. The cells were serum-starved byreplacing the growth medium with serum-free Kaighn's F-12K medium withpenicillin-streptomycin (10 U/mL) and, again, incubating overnight at37° C. and with 5% CO₂. On the third day, the medium was replaced withfresh serum-free medium and the cells were incubated with or withoutcompound (vehicle was DMSO at 0.1% maximum concentration) forapproximately 1 hour and then stimulated with 1 ng/mL recombinant humanIL-1β (R&D Systems, Minneapolis, Minn.) for 20 hours at 37° C. and with5% CO₂. Cell supernatants were collected for determination of IL-6levels using MSD (Meso Scale Discovery, Gaithersburg, Md.) 96-wellSingle Spot plates as per the manufacturer's instructions. Plates wereread with an MSD Sector Imager 6000. Prednisolone (1 μM) was used as amaximal inhibitor and defined the 100% inhibition control. Vehicle wasused to define the 0% inhibition control. Percent inhibition for eachcompound concentration, relative to these controls, was calculated usingExcel (Microsoft, Redmond, Wash.). IC50 values were generated usingGraFit 5.0 data analysis software (Erithacus Software Ltd., Surrey, UK).

TNFα IC₅₀ and % Inhibition

Human U937 pre-monocytic cells (American Type Culture Collection,Rockville, Md.) were cultured in RPMI 1640 with glutamine (2 mM),penicillin-streptomycin (10 U/mL) and 10% heat-inactivated fetal bovineserum (all from Invitrogen, Grand Island, N.Y.). Cells weredifferentiated to a monocyte/macrophage phenotype with phorbol12-myristate 13-acetate (Sigma-Aldrich, St. Louis, Mo.), 20 ng/mL,overnight. The cells were then centrifuged, the medium aspirated, thecells resuspended in an equal volume of fresh RPMI 1640 medium withglutamine and penicillin-streptomycin and fetal bovine serum as listedabove, and incubated for 48 hours at 37° C. and with 5% CO₂. Followingrecovery, the cells were scraped, counted, and plated in accordance tothe experimental design prior to stimulation with LPS, as describedbelow.

U937 cells were differentiated and plated at a density of 200,000cells/well in 96-well plates. Cells were incubated with or withoutcompound (vehicle was DMSO at 1% maximum concentration) forapproximately one hour and then stimulated with 100 ng/mLlipopolysaccharide (LPS), E. coli serotype 0111:B4 (Sigma-Aldrich, St.Louis, Mo.) for four hours at 37° C. and with 5% CO₂. Cell supernatantswere collected for determination of TNFα levels using an in-housesandwich-type ELISA. Mouse anti-human TNFα monoclonal antibody (clone28401.111) and biotinylated goat anti-human TNFα (R&D Systems,Minneapolis, Minn.) were used as the capture and detection antibodies,respectively. Streptavidin-horseradish peroxidase (HRP) (R&D Systems,Minneapolis, Minn.) and K-Blue Substrate/Red Stop (Neogen, Lexington,Ky.) were used as the detection system. Absorbance was measured at 650nm. TNFα concentrations were interpolated from a human TNFα recombinantprotein (R&D Systems, Minneapolis, Minn.) standard curve using a fourparameter logistics model by Magellan 4.11 data analysis software(Tecan, Durham, N.C.). Prednisolone (1 μM) was used as a maximalinhibitor and defined the 100% inhibition control. Vehicle was used todefine the 0% inhibition control. Percent inhibition for eachconcentration of compound, relative to these controls, was calculatedusing Excel (Microsoft, Redmond, Wash.). IC50 values were generatedusing LabStats Fit Curve V4.R7.MO data analysis software (PfizerSandwich Laboratories, UK and Tessella Support Services plc, AbingdonUK).

Ex Vivo Human Whole Blood

This study compares the inhibition of IL-1β, IFNγ, IL-6, and TNFαproduction in ex vivo LPS-stimulated human whole blood by theglucocorticoid receptor (GR) ligands Comparator A, Example 1, andprednisolone.

Venous blood from human donors was collected as 10 ml aliquots in tubescontaining sodium heparin (BD Vacutainer from Becton Dickinson andCompany, Franklin Lakes, N.Y.). Blood was added to sterile polystyreneround bottom 96-well tissue culture plates (Corning Costar) at 100μl/well, omitting the outside wells. Media (RPMI Medium 1640 withL-glutamine, Invitrogen Corporation, Carlsbad, Calif.) was added to theblood in 90 μl aliquots for a total volume of 190 μl. Outside wells werefilled with 200 μl of media. Blood was placed in a humidified 37° C.incubator with 5% CO₂ while compounds were prepared (nearly 60 minutes).

Compounds were prepared from 10 mM stock solutions in dimethylsulfoxide(DMSO, Sigma-Aldrich). Stock compound was diluted serially ⅓ in DMSO(i.e. 5 μl compound+10 μl DMSO), followed by diluting each serialdilution 1/167 into vehicle solution (2% DMSO, 30% ethanol (AAPERAlcohol and Chemical Company), and 6-8% phosphate buffered saline(Dulbecco's Phosphate Buffered Saline without calcium chloride withoutmagnesium chloride, Invitrogen Corporation, Carlsbad Calif.). Compoundor vehicle was added to blood in 10 μl aliquots as triplicates. Thefinal concentration of each prednisolone and Example 1 in the assayranged from 1000 nM to 0.457 nM. Comparator A concentrations ranged from3000 nM to 1.4 nM. Final DMSO and ethanol concentrations in the assaywere 0.1% and 1.5%. The samples were gently triturated twice to mix andreplaced in the incubator. LPS stock (E. coli serotype 0111:B4,Sigma-Aldrich), stored in aliquots of 100 μg/ml in RPMI at −20° C., wasdiluted 1/50 in RPMI to make a working stock solution. After 60 minutesof incubation, 10 μl of the prepared LPS working stock was added to theblood to a final concentration of 100 ng/ml, omitting wells to be usedas negative control. The samples were again gently triturated and theplates incubated overnight for 22 hours. Following incubation, the bloodwas centrifuged at 1500×g for 5 minutes and the plasma removed to eitherfreeze at −20° C. or assay for cytokine release.

IL-1β, IFNγ, IL-6, and TNFα protein levels were measured using MesoScale assay kits (Meso Scale Discovery, Gaithersburg, Md.). Reagentswere allowed to come to room temperature. Meso Scale plates were blockedwith 30 μl of human plasma/serum assay diluent with gentle shaking for60 minutes at room temperature. Plates were washed 3× with wash buffer(PBS, Invitrogen Corporation, with 0.05% Tween-20, Sigma-Aldrich).Calibrators for standard curves were prepared in human plasma/serumassay diluent as a ⅕ serial dilution to achieve final concentrationsranging from 50000 pg/ml to 3.2 pg/mi. Samples and calibrators wereadded at 20 μl/well, then incubated at room temperature with gentleshaking for 90 minutes. Plates were again washed 3 times with washbuffer. Detection antibody was diluted in human plasma/serum antibodydiluent to 1 μg/ml and added to the plate at 20 μl/well. Plates wereincubated as before for 60 minutes and washed again. Read Buffer T (4×)was diluted 1:1 with mqH₂O to 2 times concentration and 150 μl added toeach well. Plates were analyzed on the SECTOR Imager 6000 (Meso ScaleDiscovery) to generate raw signal values.

IL-1β, IFN-γ, IL-6, and TNFα sample values were verified to be withinthe calibrator standard curves. Individual values were compared topositive and negative controls (vehicle treated blood with LPS andvehicle treated blood without LPS, respectively) to generate %inhibition. Triplicate values were averaged for each donor. The valuesfor two donors were averaged (only one donor was used for IL-1β) andgraphed using 4-parameter fit curves in the GraFit 5.0.11 application.

Mean Values of Prednisolone Inhibition

IFNγ IL-1β Concentration (% TNFα (% IL-6 (nM) inhibition) (% inhibition)inhibition) (% inhibition) 1000 99.95181 93.85394 94.92022 62.08045333.3333 99.7687 88.98186 92.93646 40.25956 111.1111 94.99872 62.3836673.1561 11.68419 37.03704 63.51763 27.66996 37.74411 5.032463 12.3456825.733 12.86882 33.55115 −0.41737 4.115226 5.164324 5.308603 20.98221−0.23188 1.371742 10.85844 6.613491 15.1055 −0.94069 0.457247 4.9252770.525846 9.64873 2.220454

Mean Values of Example 1 Inhibition

IFNγ IL-1β Concentration (% TNFα (% IL-6 (nM) inhibition) (% inhibition)inhibition) (% inhibition) 1000 78.72981 38.06288 53.51043 8.053268333.3333 73.47381 36.04024 57.75726 2.1505 111.1111 60.63503 27.3528739.67173 0.943985 37.03704 51.01941 18.68644 38.24203 −0.82783 12.3456826.70902 9.415215 21.54167 −0.36893 4.115226 −3.18296 −1.31222 11.20262−1.06692 1.371742 19.77643 7.869405 22.38355 3.32595 0.457247 16.927238.956175 23.37486 −1.36819

Mean Values of Comparator A Inhibition

IFNγ IL-1β Concentration (% TNFα (% IL-6 (nM) inhibition) (% inhibition)inhibition) (% inhibition) 3000 28.02163 −3.03631 16.37219 −1.97032 100016.52981 −5.43701 14.96801 −0.88954 333.3333 −6.31952 −4.61436 12.23526−2.8341 111.1111 8.737671 −3.82374 8.59594 −3.49518 37.03704 −9.80677−4.19291 17.27236 −3.52461 12.34568 0.016012 0.030908 22.84851 −1.125814.115226 −1.69672 −0.86051 22.01534 −4.3436 1.371742 18.09167 18.131631.97474 1.459164

In Vivo Data

Therapeutic Therapeutic TNFα TNFα Osteocalcin Osteocalcin mCIA mCIASuppression Suppression Suppression Suppression Name (ED₅₀ dose) (ED₈₀dose) (ED₅₀ dose) (ED₈₀ dose) (ED₅₀ dose) (ED₈₀ dose) Example 1 0.4 1.50.46 1.82 2.91 >10 free base Comparator A 60 37 Comparator B 2.9 >101.64 2.91 2.39 >10 Comparator C 2.0 >10 0.27 1.33 1.01 2.84 Comparator D1.47 >20 1.18 3.06 Comparator E 0.5 2.6 0.14 1.18 0.17 1.71 ComparatorF >10 >10 2.11 5.67 1.19 >10 Comparator G 0.6 3.0 2.90 3.33 1.12 7.27Comparator H 0.23 0.72 0.51 1.46 Comparator I 17.0 >20 0.73 5.455.20 >20 Comparator J 3.0 >20 0.09 0.79 0.73 3.58 Prednisolone 1.1 5.50.90 2.10 1.00 6.80Example 1 is a potent compound in disease models.Mouse Collagen-Induced arthritis (mCIA)

Mouse Collagen-induced arthritis is a commonly used chronic, preclinicalmodel of rheumatoid arthritis in which joint swelling and bonedestruction occur following immunization with type II collagen.Reduction of disease incidence and severity has been shown previously tobe predictive of disease-modification and signs and symptoms mitigation,respectively, in a clinical setting.

In the traditional mCIA model, male DBA/J mice were immunized with 50 μgchick type II collagen (cCII) in complete Freund's adjuvant and thenboosted 21 days later with 50 μg cCII in incomplete Freund's adjuvant.Treatment with compounds was initiated on the morning of the boost andcontinued for 56 days. The effectiveness of treatment was measured bydisease incidence (i.e., number of mice showing any sign of disease) anddisease severity, both of which were measured twice a week.

In the therapeutic mCIA model, induction of disease incidence andseverity was synchronized via LPS stimulation. Male DBA/J mice wereimmunized with 100 ug of bovine type II collagen (bCII) on day 0. Allmice received an intraperitoneal injection of 20 μg of LPS on day 28 anddisease was allowed to develop through day 34. At day 34, all mice haddisease (incidence=100%) with an average severity score of seven. Dosingof compounds was initiated in the therapeutic mode on day 34 andcontinued through day 49. Different treatments were compared bymeasuring the decrease in incidence (i.e., resolution of disease) andthe decrease in severity of paw swelling over time.

Definition of mCIA Severity Scores (Maximal Score of 12/Mouse)

Severity Score Definition 1 Any redness or swelling of digits or paw 2Gross swelling of the whole paw or deformity 3 Ankylosis of jointsExample 1 had scores less than 2 for mCIA (ED₈₀) and less than 1 formCIA (ED₅₀).

TNFαand Osteocalcin (OC) Suppression

The compounds were weighed and suspended in a vehicle of 0.5%methylceuluose/0.025% tween 20 (Sigma-Aldrich, St. Louis, Mo.). Compoundsuspensions were homogenized using a Polytron PT-3100 tissue homogenizerto create a very fine suspension and were then sonicated for 10 minutesusing a water bath sonicator. Aliquots of each suspension were made fordaily dosing at 0.2 ml/dose. Swiss Webster female mice, 10-12 weeks old,28-29 grams, (Taconic, Germantown, N.Y.) were used in accordance withthe guidelines of the Institutional Animal Care and Use Committee and inaccordance with NIH guidelines on laboratory animal welfare. Mice wereacclimated in the Pfizer animal facility for three to seven days priorto being utilized in a study. Prednisolone and compounds wereadministered by oral gavage for a total of 28 days. Each treatment groupcontained 5-10 mice. To establish a dosing regimen for the studies, apilot pharmacodynamic time course experiment was conducted to quantifyTNFα repression after a single ED₈₀ dose. Compounds which suppressedTNFα were dosed QD, while compounds which did not suppress TNFα >50% outto 24 h were dosed BID.

Body weights were measured on the first and last day of each experiment.Blood samples were obtained after three weeks of dosing for asteady-state pharmacokinetic (PK) analysis. To assess compound effectson LPS-induced TNFα, all mice received an intraperitoneal injection ofLPS (Salmonella typhosa, L-7895; Sigma-Aldrich, St. Louis,) 2.5 hr afterthe last dose on day 28. Mice were sacrificed 90 minutes after LPSadministration. Serum samples were quantified for osteocalcin and TNFαusing the multiplex assays (Linco Research, Inc, St. Charles, Mo.;Luminex 100, Austin, Tx.). Samples were diluted 1:20 and the assay wasrun according to manufacturer's instructions. The osteocalcin standardwas purchased separately (Biomedical Technologies Inc., Stoughton,Mass.). Mice were fasted for 4 hours before serum was collected forTNFα, and osteocalcin levels. For each experiment, outliers weredetected by calculating the number of standard deviations from the meanof the group. If the value being examined was more than 2.5 standarddeviations from the mean, it was excluded from the rest of thecalculations.

Percent inhibition values were then calculated for each mouse using themeans of the vehicle and 10 mg/kg prednisolone control groups. Theindividual mouse percent inhibition values were fit to a four-parameterlogistic model using the dose mean for each group. Since all fourparameters were estimated and the lower plateau was not fixed at 0% andthe upper plateau was not fixed at 100%, the ED₅₀ and ED₈₀ values werecalculated by using an inverse calibration formula for a response equalto 50% or 80% inhibition or activation.

ED₅₀ and ED₈₀ values are the doses (in mg/kg) required to result in a50% or 80% effect, respectively, on a particular endpoint. ED₅₀ and ED₈₀values were obtained for the various endpoints using a four-parameterlogistic fit. For compounds which were tested multiple times, ED₅₀ andED₈₀ values were obtained using four-parameter logistic fits of thecombined data from multiple experiments. For compounds which did notachieve an 80% effect, the ED₈₀ value is designated as >10 mg/kg or >20mg/kg, depending on the highest dose tested.

House Dust Mite Model of Asthma

Mice were treated with three doses of Example 1 (0.1, 1 and 10 mg/kg,p.o., b.i.d.) or prednisolone (0.1, 1 and 10 mg/kg, p.o., b.i.d.).Separate groups of animals were treated with the respective vehicles,and no effect inflammatory cell influx into house dust mite-induced BALinflammatory cell influx was demonstrated.

Example 1 mitigated cell infiltration into BAL fluid dose-dependently.Evaluation of BAL fluid cell types using flow cytometry showedsignificant reductions in eosinophils, neutrophils, lymphocytes andT-cells. In comparison, prednisolone conferred similar reductions in BALfluid cell infiltration at similar doses (data not shown).

Lympho- Exam- Total cells Eosinophils Neutrophils cytes T-cells ple 1 (%(% (% (% (% (mg/kg) inhibition) inhibition) inhibition) inhibition)inhibition) 0.1 22 42 29 34 39 1 72 97 97 94 88 10 83 99 100 97 96

Dissociation Index

A dissociation index (DI) was chosen as a measurement to quantify thedissociation of compounds relative to that of prednisolone in terms ofbiomarkers of anti-inflammatory efficacy and side-effects. Dissociationindices were calculated using clinically relevant biomarkers that couldbe utilized in early clinical development. Serum osteocalcin andLPS-induced serum TNFα are accepted clinically as predictive for boneformation and anti-inflammatory efficacy, respectively.

The dissociation index was based on the following tenets:

-   1) Dissociation required a dose-margin between biomarkers of    inflammation and side-effects and was defined by the formula:

${D\; I} = \frac{{Side}\text{-}{effect}\mspace{14mu} {endpoint}}{{Anti}\text{-}{inflammatory}\mspace{14mu} {endpoint}}$

For example:

${D\; I} = {\frac{{osteocalcin}\mspace{14mu} {supprepression}\mspace{14mu} ({OC})\mspace{11mu} {ED}_{50}\mspace{11mu} ( {{or}\mspace{14mu} {EAUC}_{50}} )}{{TNF}\; \alpha \mspace{14mu} {supprepression}\mspace{14mu} ( {{TNF}\; \alpha} )\mspace{11mu} {ED}_{50}\mspace{11mu} ( {{or}\mspace{14mu} {EAUC}_{50}} )}.}$

-   2) The DI of a compound can be considered relative to that observed    with prednisolone, its clinical comparator. The corrected or    normalized DI was defined as compound DI divided by prednisolone DI.

Dissociation Index (ED₅₀ and ED₈₀)

NAME OC/TNFα (ED₅₀) OC/TNFα (ED₈₀) Example 1 free base 6.33 >5.49Comparator A 0.62 Comparator B 1.46 >3.44 Comparator C 3.74 2.14Comparator D 0.80 <0.15 Comparator E 1.21 1.45 Comparator F 0.56 >1.76Comparator G 0.39 2.18 Comparator H 2.22 2.03 Comparator I 7.12 >3.67Comparator J 8.11 4.53 Prednisolone 1.11 3.24

Example 1, Comparator I, Comparator J had a DI greater than 5 forOC/TNFα (ED₅₀). Example 1 and Comparator J had a DI greater than 4 forOC/TNFα (ED₈₀).

Corrected Dissociation Index (ED₅₀ and ED₈₀) Based on Prednisolone

Name OC/TNFα (ED₅₀) OC/TNFα (ED₈₀) Example 1 free base 5.70 >1.69Comparator A 0.56 Comparator B 1.32 >1.06 Comparator C 3.37 0.66Comparator D 0.73 <0.05 Comparator E 1.09 0.45 Comparator F 0.50 >0.54Comparator G 0.35 0.67 Comparator H 2.00 0.63 Comparator I 6.41 >1.13Comparator J 7.31 1.40

Example 1, Comparator I, Comparator J had a corrected DI greater than 5for OC/TNFα (ED₅₀). Example 1 had a corrected DI greater than 1.50 forOC/TNFα (ED₈₀).

EAUC₅₀ and EAUC₈₀

Drug exposure, defined as drug plasma concentrations integrated overtime (AUC), was used to make pharmacodynamic comparisons betweenprednisolone and Example 1. Due to the short half-lives of prednisoloneand Example 1 in the mouse, AUC (0-4 hr) values accounted for more than95% of the AUC (0-24 hr) values. Due to blood volume samplinglimitations in mice, AUC (0-4 hr) values were used to makepharmacodynamic comparisons.

Prednisolone Example 1 (AUC 0-4 hr) (AUC 0-4 hr) (μg * hr/mL) (μg *hr/mL) EAUC₅₀ EAUC₈₀ EAUC₅₀ EAUC₈₀ Serum TNFα 0.81 0.95 0.22 0.33 Serumosteocalcin 0.56 1.60 1.30 4.20 Cortical bone 0.09 0.60 0.27 1.55formation rate (BFR) Disease incidence 0.25 0.60 0.04 0.10 (traditionalmCIA) Disease severity 0.38 1.19 0.09 0.19 (therapeutic mCIA)Dissociation Index OC/TNFα^(a) 0.7 1.7 5.9 12.7 BFR/TNFα^(a) 0.1 0.6 1.24.7 BFR/disease 0.4 1.0 6.8 15.5 incidence^(b) BFR/disease 0.2 0.5 3.08.2 severity^(b) Corrected Dissociation Index OC/TNFα^(a) 8.4 7.5BFR/TNFα^(a) 12.0 7.8 BFR/disease 17.0 15.5 incidence^(b) BFR/disease15.0 16.4 severity^(b) ^(a)Data compared in mice in the same model.^(b)Data compared across mice in different models using plasma exposureto normalize. BFR, bone formation rate.

EAUC₅₀ and EAUC₈₀ Data Set 2

Example 1 Prednisolone Parameter EAUC₅₀ EAUC₈₀ EAUC₅₀ EAUC₈₀ 28-dayrepeat dose model Serum TNFα 0.22 0.33 0.82 0.95 Serum osteocalcin (OC)1.31 4.15 0.56 1.64 Cortical bone formation 0.35 1.76 0.10 0.66 rate(BFR) Disease incidence — 0.10 — 0.50 (traditional mCIA) Diseaseseverity 0.09 0.19 0.38 1.19 (therapeutic mCIA) Disease incidence 0.120.33 0.68 1.38 (therapeutic mCIA) Corrected dissociation indices (DI)OC/TNFα 8.6 7.4 BFR/TNFα 16.0 7.6 BFR/disease incidence 14.5 10.6BFR/disease severity 13.0 15.5Example 1 is a dissociated compounds Example 1 had a DI and a correctedDI for EAUC₅₀ and EAUC₈₀ greater than 7 for OC/TNFα, BFR/TNFα,BFR/disease incidence, and BFR/disease severity.

Cortical Bone Histomorphometry to Determine BFR

During the in-life portion of each study, mice received twointraperitoneal (i.p.) injections (20 mg/kg, 100 ml/mouse) of calcein(C-0875; Sigma-Aldrich, St. Louis, Mo.) on days 1 and 26 for bonehistomorphometry measurements. Calcein incorporates into the bonemineral and allows measurement of bone formation rate. Calcein wasdissolved in 2% sodium bicarbonate. During the tissue harvest, the lefttibiae were excised and cleaned for cortical histomorphometrymeasurements. After all skin and muscle were removed, tibiae were placedin 70% ethanol (4° C.) in the dark for a minimum of 24 hours.

Ground transverse sections were used for histomorphometric analysis ofcortical bone. Bones were sectioned using a low-speed saw (Isomet,Buehler, Lake Bluff, Ill.) equipped with a diamond wafer blade. The endof each tibia was removed proximal to tibia-fibula synostosis and a 75mm cross-section was cut. Using a roughened glass plate and a cork,sections were ground to ˜25 mm until transparent and all labels weredistinguishable under a fluorescent microscope. Sections were dehydratedusing the following solutions for a minimum of two minutes each: 1) 70%ethanol, 2) 95% ethanol, 3) 100% ethanol, 4) 50/50 ethanol/xylene, and5) xylene (twice) (#534056; Sigma-Aldrich, St. Louis, Mo.). Sectionswere mounted using Eukitt Quick Mounting Medium (#03989, Sigma-Aldrich,St. Louis, Mo.) after which coverslip was applied. Using theOsteomeasure Bone Analysis Program (Osteometrics, Inc., Decatur, Ga.),bone formation rate was calculated by tracing the 1^(st) and 3^(rd)fluorescent labels and the inner and outer perimeter of the bone. Boneformation rate was calculated by the following equation: (Inter-labelwidth/Label interval)*(Labeled perimeter/Bone perimeter). At least fivesamples were measured from each treatment group in each study.

1. A method comprising contacting a glucocorticoid receptor with thecompound of Formula I:

wherein R¹ is —H; or salt thereof.
 2. A method of treating a conditionmediated by glucocorticoid receptor activity in a subject comprisingadministering to the subject a compound of Formula I:

wherein R¹ is —H; or salt thereof.
 3. The method of claim 7, wherein thecondition is an inflammation related condition.
 4. The method of claim7, wherein the condition is asthma, dermatitis, inflammatory boweldisease, Alzheimer's disease, psychotic major depression, neuropathy,transplant rejection, multiple sclerosis, chronic uveitis, or chronicobstructive pulmonary disease.
 5. The method of claim 7, wherein thecondition is rheumatoid arthritis.
 6. The method of claim 7, wherein thecondition is dermatitis.
 7. The method of claim 7, wherein the conditionis asthma.
 8. The method of claim 7, wherein the condition isAlzheimer's disease.